SYNDECAN STIMULATION OF CELLULAR DIFFERENTIATION

Abstract: Methods are provided for altering levels of syndecan within a cell. The methods include enhancing syndecan expression via administration of growth factors, preventing suppression of syndecan expression via administration of anti-steroid agents, and altering syndecan biochemistry within the cell. The methods are used to induce or maintain cellular differentiation, and to decrease the growth of malignant cells. Application of the methods to the treatment of patients, including humans, is provided.

Description: SYNDECAN STIMULATION OF CELLULAR DIFFERENTIATION FIELD OF THE INVENTION This invention is in the field of cancer biology and therapy. Specifically, the invention is to a method for altering the differentiated state of a cell by altering syndecan expression. The method allows for the normalization of the growth rate and differentiation state of malignant cells, such method being based on the stimulation of syndecan expression in malignant cancer cells. Re- expression of syndecan in such malignant cells promotes their normal differentiated phenotype and prevents their tumor formation. This method may also be applied to normal cells to increase their differentiation, and therefore support the maintenance of cells, e.g. keratin production to prevent baldness. BACKGROUND OF THE INVENTION It is becoming more evident that cell surface proteoglycans play an important role in the regulation of cell behavior (Ruoslahti etal., Cell 64:867- 869 (1991)). Through their covalently bound glycosaminoglycan side chains, such proteoglycans can bind various extracellular effector molecules (Jalkanen, βtal., in Receptors for Extracellular Matrix, J. MacDonald & R. Mecham, Editors, Academic Press, San Diego, pp. 1-37 (1991)). One central challenge in protθoglycan biology is the need to understand what directive a cell receives by binding different effector molecules via the cell surface proteoglycans. It is further imperative to investigate what kind of intracellular response such binding activates, thereby leading to altered behavior of the cell. The way to approach these questions is to create biological models which are dependent on the expression of any given proteoglycans. Syndecan is the best characterized cell surface protθoglycan (Saunders era/., J. Cell Biol. 703:1547-1556 (1989); Mali etal., J. Biol. Chem. 265:6884- 6889 (1990)). It was originally isolated from mouse mammary epithelial (NMuMG) cells as a hybrid proteoglycan containing both heparin sulfate and chondroitin sulfate glycosaminoglycan side chains (Rapraeger et al., J. Biol. Chem. 260:11046-11052 (1985)). Recent studies have revealed its expression, not only on epithelial cells but also on differentiating fibroblasts of developing tooth (Thesleff etal., Dev. Biol. 129:565-572 (1988); Vainio etal., J. Cell Biol. 705:1945-1964 (1989)), on endothelial cells of sprouting capillaries (Elenius βtal., J. Cell Biol. 114:585-596 (1991)) and on the surface of lymphocyte subpopulations (Sanderson etal., Cell Regul. 7:27-35 (1989)) intimating that its function can vary on the surfaces of different cells. Syndecan belongs to a family of proteoglycans with conserved plasma membrane and cytoplasmic domains but with dissimilar ectodomains (Mali et al., J. Biol. Chem. 265:6884- 6889 (1990)). The conserved structure of syndecan suggests that it could participate in signal transduction through the plasma membrane. Syndecan binds several extracellular effector molecules but this binding is selective. For example, syndecan binds interstitial collagens and fibronectin but does not bind to vitronectin or laminin (Koda et al., J. Biol. Chem. 260:8156-8162 (1985)); Saunders er a/., J. Cell Biol. 706:423-430 (1988); Elenius et al., J. Biol. Chem. 265:17837-17843 (1990)). Moreover, syndecan isolated from tooth mesenchyme has revealed selective binding to tenascin not observed for syndecan from NMuMG cells (Salmivirta et al., J. Biol. Chem. 266:7733-7739 (1991)). This suggests variation in syndecan glycosylation that results in the selective binding properties for syndecan. Polymorphism of syndecan glycosylation has also been observed between simple and stratified epithelia (Sanderson er a/., Proc. Natl. Acad. Sci. USA 55:9562-9566 (1988)); but whether these changes also reflect altered ligand recognition by syndecan remains unknown. Syndecan also binds growth factors, such as basic fibroblast growth factor (bFGF) (Kiefer etal., Proc. Natl. Acad. Sci. USA 67:6985-6989 (1990); Elenius et al., J. Biol. Chem. 267:6435-6441 (1992)). Very recently, Yayon and coworkers (Yayon etal., Cell 64:841-848 (1991)) and Rapraeger and co-workers (Rapraeger etal., Science 252:1705-1708 (1991)) have shown that hepar in-like molecules are required for the binding of bFGF to its high affinity receptor, indicating that syndecan-like molecules can also modulate the growth factor response. The fact that cell surface proteoglycans can bind both growth factors and matrix components could theoretically imply a role in regulating, both temporally (timing of expression) and spatially (precise localization), growth promotion by immobilizing these effector molecules to the vicinity of cell-matrix interactions. This is supported by the intriguing pattern of syndecan expression in the development that follows morphogenetic rather than histological patterns (Thesleff et al., Dev. Biol. 129:565-572 (1988); Vainio et al., J. Cell Biol. 705:1945-1954 (1989) and Vainio et al., Dev. Biol. 734:382- 391 (1989)), and moreover, that syndecan expression is localized to sites of active proliferation (Elenius et al., J. Cell Biol. 774:585-596 (1991) and Vainio et al., Dev. Biol. 747:322-333 (1991)). In simple epithelium, syndecan is polarized to baso-lateral surfaces where it co-localizes with actin rich cytofilaments (Rapraeger et al., J. Cell Biol. 703:3683-2696 (1986)). Upon rounding, syndecan is shed from the cell surface by proteolytic cleavage of the core protein at the cell surface, a process which separates the matrix binding ectodomain from the membrane domain (Jalkanen etal., J. Cell Biol. 705:3087-3096 (1987)). By this way, syndecan has been proposed to be involved in the maintenance of epithelial morphology. Mouse mammary tumor cells (S115), when steroid-induced to modulate their morphology from an epithelial to a more fibroblastic or fusiform phenotype, lose syndecan expression (Leppa etal., Cell Regul. 2:1-11 (1991)), as to several other cell types in vivo when they transform (Inki et al., Am. J. Pathol. 739:1333-1340 (1991); Inki et al., Lab. Invest. 66:314-323 (1992)), suggesting a general loss of syndecan expression during malignant transformation. SUMMARY OF THE INVENTION It is therefore an object of the subject invention to provide a method of altering the differentiated state of a host cell by altering syndecan expression in such cell. It is a further object of the invention to provide a method to induce and regulate syndecan expression, especially in cells which exhibit a malignant phenotype, regardless of the origin of transformation. It is a further object of the invention to provide a treatment for the reduction of tumor growth in a patient in need of such treatment, by administration of a composition to such patient, such composition comprising efficacious amounts of one or more agents that stimulate syndecan synthesis in the tumor cells of such patient. It is a further object of the invention to provide the DNA sequence and localization of promoter, suppressor and enhancer elements for the syndecan gene. It is a further object of the invention to provide a method for the enhancement of syndecan expression in a host cell, by enhancing syndecan gene transcription. It is a further object of the invention to provide a method for the enhancement of syndecan expression in malignant cells, by preventing suppression of syndecan gene transcription. It is a further object of the invention to provide a biochemical method for the inactivation of suppressors of syndecan gene expression in malignant cells. It is a further object of the invention to provide a method for the stimulation of cellular differentiation by enhancing syndecan expression in both malignant and normal cells. It is a further object of the invention to provide a method for the stimulation of cellular proliferation and differentiation, thus tissue regeneration, especially in processes such as wound healing, by enhancing syndecan expression. Further features, objects and advantages of the present invention will become more fully apparent from a detailed consideration of the following description of the subject invention when taken together with the accompanying drawings. BRIEF DESCRIPTION Q THE DRAWINGS Figure 1. Figure 1 is a diagram of the assembly of mouse syndecan gene and its promoter region. Figure 2. Figure 2 is the complete sequence of the mouse syndecan gene [SEQ ID Nos. :1 : (DNA) and :2: (protein)] and its proximal promoter. Regulatory sites for the expression of syndecan may also exist on the first intron following the first exon (see Figure 1). Figure 3. Figure 3 is a diagram of the assembly of mouse syndecan promoter region and the localization of the enhancer and suppressor elements together with restrictions sites for three different enzymes. Figure 4. Figure 4 is the complete sequence of the mouse syndecan enhancer element [SEQ ID No. :3: (DNA)].located 8-10 kbs upstream from the transcription initiation site as indicated in figure3. Figure 5. Figure 5A (panels A-D) is a photographic presentation of reduced growth ability of syndecan-transfected cells in soft agar. Panel A (wild- type S115 cells) and B (control transfected cells) are pictures of the colonies that are formed in soft agar in the presence of testosterone, a feature typical for hormone-transformed cells. This growth ability was not observed with two independent syndecan-transfected cell clones (panels C and D), demonstrating how syndecan re-expression can overcome the effect of hormone-induced transformation. Figure 5B is a graphical presentation of how syndecan- transfected cells lose their ability to form tumors in nude mice. Wild-type or control transfected cells produce tumors in testosterone-administered nude mice while syndecan transfected cells revealed a very low tendency to produce tumors. Figure 6 is a graphical representation of enhanced syndecan expression in 3T3 cells by simultaneously administered basic fibroblast growth factor (bFGF) and transforming growth factor beta (TGF-β). This is an example of how syndecan expression can be enhanced as a result of growth factor action in normal cells during the differentiation process. Figure 7 is a graphical representation of enhanced syndecan expression by MCF-7 cells exposed to the anti-estrogen toremifene. When exposed to estrogen, syndecan expression in MCF-7 cells was reduced and the cells transformed. Subsequent treatment with the anti-estrogen (toremifene) restored syndecan expression to levels close to that found in cells not exposed to estrogen and aided the cells in maintaining their normal growth behavior. Figure 8 is a graphical presentation of how the suppressor element (see figure 3) is acitve in S115 cells treated with testosterone. Indicated stretches of promoter sequences were transfected in hormone-treated S115 cells and analysed for their transcription activity as described in Example VI. A dramatic drop was observed wiht suppressor construct as indicated in Figure 3, and it was more obvious in transformed S 115 (a) cells than in 3T3 cells (b). Figure 9 is a graphical presentation of how the enhancer element is active in growth hormone-treated 3T3 cells. Various stretches of promoter were transfected in 3T3 cells and analysed for their transcription activities. Fragment pXb6, which is the same as illustrated in Figure 3 as an enhancer, revealed more than ten fold stimulation in 3T3 cells exposed to growth factors bFGF and TGFβ if compared to non-treated cells. DEFINITIONS In order to provide a clearer and more consistent understanding of the specification and claims, including the scope to be given such terms, the following definitions are provided. "Enhancement" or "Stimulation" of Syndecan expression. By "enhancement" or "stimulation" of syndecan expression" is meant an effect of increasing the synthesis of syndecan, either by the induction or de-suppression (de-repression) of syndecan gene transcription and/or translation. Cell growth. By "cell growth" is meant cell replication, both controlled and uncontrolled. Malignant. By "malignant" is meant uncontrolled cell growth. More Differentiated Phenotype. In stating that a cell has a "more differentiated phenotype" is meant that the cell possesses a phenotype usually possessed by a certain cell type more differentiated than the cell, which the cell was deficient in prior to enhancement of syndecan expression according to the invention. This phenotype may be defined by one or more phenotypic characteristics. For example, an epithelial cell is a more differentiated phenotype of a mesenchymal-like shape; therefore, the ability of the method of the invention to maintain cells in an epithelial cell morphology rather than a mesenchymal-like shape is a more differentiated phenotype within the meaning of the definition. Continuous syndecan expression is necessary for the maintenance of terminal differentiation of epithelial cells. Syndecan expression is also linked to the normal differentiation of mesenchymal cells. However, only a transient increase in syndecan expression is needed for normal differentiation in mesenchymal cells. Therefore, contrary to epithelial cells, expression of syndecan is not needed for maintenance of terminal differentiation in mesenchymal cell. To induce differentiation of a suitable mesenchymal precursor cell population (such as a "condensing mesenchymal" cell population) to a fully differentiated mesenchymal cell, there is needed only a transient expression of syndecan expression. Therefore, a terminally differentiated mesenchymal cell is a "more differentiated phenotype" than a condensing mesenchymal cell. Other useful phenotypes that are present in syndecan-deficient cells and not in cells expressing sufficient syndecan include fusiform shapes with long filopodial extensions with extensive under-and overlapping of these processes (so that the cells appear to have a defect in cell adhesion). in another example, syndecan-deficient NMuMG cells continue to secrete milk fat globule antigen (and thus appear mammary-like) and continue to express cytokeratins (thus appear epithelial-like). However, their actin- containing cytoskeleton is disorganized and their expression of beta-j integrins and E-cadherins at the cell surface is markedly reduced. Upon expression of sufficient syndecan, these phenotypes are corrected so that there is no reduction in cell surface integrins or E-cadherin and the cell has an epithelial morphology. Therefore, the amount of cell surface integrins or E-cadherin are useful markers of syndecan expression and may be used to monitor what amount of a drug is needed for efficacious results according to the method of the invention. Efficacious Amount. An "efficacious amount" of an agent is an amount of such agent that is sufficient to bring about a desired result, especially upon administration to an animal or human. Administration. The term "administration" is meant to include introduction of agents that induce syndecan expression into an animal or human by any appropriate means known to the medical art, including, but not limited to, injection, oral, enteral and parenteral (e.g., intravenous) administration. Pharmaceutically Acceptable Salt. The term "pharmaceutically acceptable salt" is intended to include salts of the syndecan-inducing agents of the invention. Such salts can be formed from pharmaceutically acceptable acids or bases, such as, for example, acids such as sulfuric, hydrochloric, nitric, phosphoric, etc., or bases such as alkali or alkaline earth metal hydroxides, ammonium hydroxides, alkyl ammonium hydroxides, etc. Pharmaceutically Acceptable Vehicle. The term "pharmaceutically acceptable vehicle" is intended to include solvents, carriers, diluents, and the like, which are utilized as additives to preparations of the syndecan-inducing agents of the invention so as to provide a carrier or adjuvant for the administration of such compounds. Treatment. The term "treatment" or "treating" is intended to include the administration of compositions comprising efficacious amounts of syndecan- inducing agents to a subject for purposes which may include prophylaxis, amelioration, prevention or cure of a medical disorder, including tumor growth, hair growth. ~ DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS Briefly, in its broader aspects the present invention comprehends a method for maintaining a differentiated phenotype in a normal (non-malignant) cell that otherwise would suppress syndecan expression, by maintaining syndecan expression in such cell. The invention also comprehends a method for inducing a more differentiated phenotype in a malignant cell that lacks (or is deficient in) syndecan expression, by stimulating syndecan expression in such a cell. As used herein, if a cell is said to "lack" syndecan expression, it is meant to include cells that are either completely deficient in syndecan protein or produce insufficient syndecan levels to maintain or attain a desired differentiated phenotype. The methods of the invention will not only prevent the progression (worsening) of a transformation state and tumor growth of cells, but can also be used to maintain differentiated cells in their differentiated state so that they continue to perform differentiated functions characteristic of such cells. Examples of differentiated functions of non-malignant cells include the secretion characteristics of a cell (that is, the secretion of specific proteins and/or other macromolecules) and hair formation by epidermal cells of skin. Thus, according to the invention, administration of agents capable of inducing syndecan expression in epidermal skin cells of the scalp will promote hair growth among bald (or balding) people. The subject method may be accomplished in a variety of manners including biochemical, chemical or even molecular biological type methods. While the method is applicable to a variety of cancer (both malignant and non- malignant) and normal cells, it is particularly adaptable for treating malignant cells which have become transformed. This includes cells transformed due to hormonal influences of the body or environmental influences, such as chemicals or radiation exposure. Especially, the tumor type is a tumor characterized by loss of syndecan, for example, a glioma, myeloma, carcinoma, sarcoma, lymphoma, or adenoma. Generally, any cell genetically capable of expressing syndecan can be a cell stimulated to express syndecan by the method of the invention. Syndecan is naturally expressed in a wide variety of epithelial cells in mature and embryonic tissues and by various embryonic mesenchymal tissues undergoing inductive interactions with epithelia. In addition, syndecan is naturally expressed on Leydig cells and on developing B-lymphocytes and a subpopulation of plasma cells. Enhanced syndecan expression may be achieved by administration of compositions containing a biochemically, and/or chemically and/or molecular- biologically active component to an individual. Compositions may be administered orally, intravenously, subcutaneously or locally, or by any other method which will allow cells, normal or malignant, to be exposed to syndecan expression enhancing component. By a "biochemically" or "chemically" active component is meant a component that alters the endogenous syndecan biochemistry or chemistry of the target cell without altering syndecan gene expression per se. For example, such alterations may include altering the half-life of syndecan protein or mRNA, so as to increase levels of syndecan protein in the cell, for example, by altering the external domain of the cell's endogenous syndecan, or the cell surface membrane properties in general, so as to retain higher levels of syndecan on the cell surface; and, altering the syndecan protein active site(s), so as to enhance the efficiency of the syndecan response. By a "moiecular-biologically" active component is meant a component that alters endogenous syndecan gene expression in a manner that allows for an increase in cellular syndecan, such as, for example, by stimulating transcription, preventing (or reducing) suppression of transcription, de- repression transcription, and generally increasing levels of mRNA and/or translation efficiency. It is known that cellular transformation involves activation of some cell growth stimulating genes (like oncogenes) and inactivation of some other genes, which work to suppress cell growth. It has recently been shown that loss of syndecan expression is observed upon transformation of cells, and that this suppression is due to syndecan gene inactivation (Leppa et al., Cell Regul. 2:1- 11 (1991); Inki etal., Am. J. Pathol. 139:1333-1340 (1991); Inki etal., Lab. Invest. 66:314-323 (1992)). This was demonstrated in several biological models of various known carcinogenic systems (oncogenes, chemical carcinogens, UV-light, hormone-exposure, etc.). Therefore, syndecan gene suppression is implicated in the development of cellular transformation. Further, according to the invention, all the manipulations of such cells which can induce syndecan expression in malignant cells, also induce these cells to obtain a more differentiated phenotype, and thus, subsequently reduce their potential tumorigenic behavior and metastasis. In a preferred embodiment, the cell in which syndecan expression is stimulated is a cell that is steroid-responsive. Examples of such steroid- responsive cells include breast cells, endometrium cells and prostate cells, especially in the malignant state. In a highly preferred embodiment, the cell is responsive to estrogen and/or androgen. Examples of other cell types that will respond to the treatment of the invention include malignant and non-malignant mesenchymal cells. The regulatory elements of a given gene are commonly located upstream from (5' to) the transcription initiation site. Syndecan, however, has a very peculiar gene structure, in which the first and second exons are separated by a very large intron (Figure 1). This could mean that, in addition to the base sequences upstream from the transcription site, syndecan expression may also be susceptible to regulation by base sequences located between first and second exons (Figure 2), that is, in the first intron. It has now surprisingly been found that the syndecan gene has a strong enhancer element located 8-10 kb upstream from the transcription initiation site. The sequence of this enhancer element has been identified and is given in Figure 4 [SEQ ID. No. 3]. Manipulation of the upstream region of the syndecan gene can block its inactivation during malignant transformation. For example, replacement of the region in front of first exon of the syndecan gene with the glucocorticoid- inducibie elements of mouse mammary tumor virus (MMTV) not only blocks syndecan suppression during malignant transformation, but also inhibits the ability or potential of cells to transform and become tumorigenic (Figure 3). These findings suggest a very important role for syndecan in the maintenance of normal epithelial morphology (Leppa et at., Proc. Natl. Acad. Sci. USA 59:932-936 (1992)). Cells destined to differentiate during organ formation or tissue regeneration also exhibit enhanced syndecan expression (Vainio etal., Dev. Biol. 747:322-333 (1991); Elenius etal., J. Cell Biol. 774:585-595 (1991)). The component(s) responsible for the regulation of syndecan expression (either directly or indirectly) have not yet been identified. Growth factors are candidates for this role tince they are known to be involved in the regulation of early development and cellular differentiation (Heath et al., Curr. Opin. Cell Biol. 3:935-938 (1991)). Involvement of growth factors is also supported indirectly by the fact that the expression of two embryonally important growth factors (TGF-β and FGF) has been shown to coincide with syndecan within developing tooth (Vaahtokari etal., Development 773:985-994 (1991); Wilkinson etal., Development 705:131-136 (1989)). Based on these findings the possible effect of growth factors on the expression of syndecan has been tested. It was shown that both bFGF and TGF-β alone, but especially if administered in combination, enhance syndecan expression by 3T3 cells (Figure 4). This stimulation is close to the levels observed in syndecan-expressing epithelial cells (Elenius etal., J. Biol. Chem. 267:6435-6441 (1992)) prior to their malignization (Leppa et al., Proc. Natl. Acad. Sci. USA 59:932-936 (1992)). These findings suggest that growth factors, and their derived fragments and domains may prove to be valuable for the development of an active tool for the regulation of syndecan expression. Preferably, for treatment of humans and animals, a drug is administered that results in the enhancement of syndecan expression to levels sufficient to facilitate cellular differentiation in the degenerative stages of tissues. Such drugs are herein termed "syndecan-inducing agents." Syndecan-inducing agents include the growth factors and their derivatives that retain growth-factor activity. Examples of such growth factors include bFGF, and TGF-β, whether administered separately or together. Even more preferred is a syndecan-inducing agent that has good tissue and cell penetration so that it could directly interfere with suppressor(s) of syndecan expression within cell nuclei. Such a syndecan-inducing agent is the known antitumor drug toremifene. When toremifene, known to have good plasma membrane penetration, is administered to the hormone-transformed epithelial cells with reduced syndecan expression, the cells reverse their lowered syndecan expression, and induce syndecan levels to those close to the level observed with normal, non-transformed cells (Figure 5). This demonstrates that syndecan-inducing agents useful in the methods of the invention are known and available and that such agents can specifically prevent cellular malignization by blocking suppression of syndecan expression. Another useful drug in this regard is tamoxifen. Such syndecan-inducing agents may be administered using currently available preparations, or in any pharmaceutically acceptable vehicle. The route of administration may be any route that delivers efficacious levels of the drug to the desired active site, for example, by injection. For parenteral administration, preparations containing one or more syndecan-inducing agents may be provided to the patient in need of such treatment in combination with pharmaceutically acceptable sterile aqueous or non-aqueous solvents, suspensions and emulsions. Examples of non-aqueous solvents are propylene glycol, polyethylene glycol, vegetable oil, fish oil, and injectable organic esters. Aqueous carriers include water, water-alcohol solutions, emulsions or suspensions, including saline and buffered medical parenteral vehicles including sodium chloride solution, Ringer's dextrose solution, dextrose plus sodium chloride solution, Ringer's solution containing lactose, or fixed oils. Intravenous vehicles include fluid and nutrient replenishers, electrolyte replenishers, such as those based upon Ringer's dextrose and the like. The syndecan-inducing agent of the invention can be employed in dosage forms such as tablets, capsules, powder packets, or liquid solutions for oral administration if the biological activity of the syndecan-inducing agent is not destroyed by the digestive process and if the characteristics of the compound allow it to be absorbed across the intestinal tissue. The syndecan-inducing agents may also be administered by means of pumps, or in sustained-release form. The syndecan-inducing agents used in the method of invention may also be delivered to specific organs in high concentration by means of suitably inserted catheters, or by providing such molecules as a part of a chimeric molecule (or complex) which is designed to target specific organs. Administration in a sustained-release form is more convenient for the patient when repeated injections for prolonged periods of time are indicated. The composition containing the syndecan-inducing agent can be manufactured in a manner which is in itself know, for example, by means of conventional mixing, granulating, dragee-making, dissolving, lyophilizing or similar processes. The compositions of the present invention that provide the syndecan-inducing agent, in and of themselves, find utility in their ability to slow or prevent tumor growth or tumor reappearance. The syndecan-inducing compositions of the invention utilize the body's own mechanisms for promoting differentiation of specific cell types to its maximum potential. In intravenous dosage form, the compositions of the present invention have a sufficiently rapid onset of action to be useful in the acute management of tumor growth. Additionally, a low potency version is useful in the management of disorders wherein a tumor has been effectively treated and the patient appears to be in remission, but it is desired to maintain sufficient levels of syndecan- inducing agents in the patient so as to assist the body in preventing a recurrence of the tumor. Typical doses of toremifene or tamoxifen, and other such syndecan- inducing agents useful in the methods of the invention for treatment of humans or other animals are 20-600 mg daily, and preferably 20-60 mg daily. The examples below are for illustrative purposes only and are not deemed to limit the scope of the invention. j= M£LEi Reversal of hormone-induced transformation by exogenous syndecan expression. As previously described (Leppa etal., Cell Regulation 2:1-11 (1991)), S115 mouse mammary tumor cells were routinely cultured in DMEM. For experimental studies involving hormone treatment, inactivated fetal calf serum (i-FCS) was replaced with 4% dextran charcoal-treated fetal calf serum (DCC- FCS), which eliminates endogenous steroids from serum), and used either with or without testosterone (10 nM) and with or without dexamethasone (10 nM or 1 μM). Cells were plated at a density of 10,000 cells/cm 2 and the medium was replenished every 3 days. Plasmid pUC19-hsynpr7 containing human syndecan cDNA (Mali etal., J. Biol. Chem. 265:6884-6889 (1990)) was digested with Nae\ restriction endonuclease, and the derived 336 bp long-fragment was separated in and eluted from low melting agarose gel. Plasmid pUC19-hsyn4 (Mali etal., J. Biol. Chem. 265:6884-6889 (1990)) was digested with Nae\ and Hindtt (polylinker site), and the plasmid-containing fragment starting from base 487 was isolated. The Nae\ fragment from hsynpr7 was ligated to the pUC-hsyn Nae\/Hindl\ digested vector. The orientation of insert was verified by restriction enzyme analysis and sequencing. The derived plasmid, containing the full coding region of human syndecan core protein, was named pUC19-hsynfull. This plasmid was further digested with Bam\λ\ and Sph\ (poiylinker site). A fragment containing syndecan coding region bases 150-1461 was isolated and blunt- ended, using Klenow and T4 DNA polymerase. Finally, this fragment was ligated to Sa/l-linearized and blunt-ended pMAMneo vector (Clontech; Palo Alto, CA), resulting in a chimeric gene containing RSV-MMTV-LTR promoter connected to human syndecan coding region and SV-40 polyadenylation signal. The orientation was confirmed by restriction enzyme digestions. The plasmid was named pMAMneo-hsyn. For control transfections, a 642 bp long-H/ndlll/Piα/ll fragment of human growth hormone gene (consisting of exons 4 and 5; Bornstein etal., J. Biol. Chem. 263:1603-1606 (1988)) was blunt-ended and cloned into the same pMAMneo vector, as described above. This control construct was named pMAMneo-hGH. All plasmids were isolated using the CsCI density gradient method. Before transfections, plasmids were linearized with MliΛ, chloroform/phenol extracted and ethanol precipitated. Transfections were performed using Lipofectin™ (BRL), according to manufacturer's instructions. After selection for two weeks (G418; 750 μg/ml, Sigma), surviving clones were isolated from growth plates using cloning cylinders. The expression of human syndecan or growth hormone (consisting of exons 4 and 5) mRNAs was then confirmed by RNA isolation and Northern blot analysis. Clones expressing high levels of transfected genes were selected for further studies and characterizations. These stock cells were routinely cultured in the presence of G418 (300 μg/ml). For the measurement of exogenous syndecan expression total RNA was isolated from wild-type S115 cells and cells transfected with human syndecan or growth hormone genes. RNA was extracted using 4M guanidine isothiocyanate and CsCI pelleting, as earlier described by Chirgwin et al., Biochemistry 73:5294-5299 (1979)). RNA from normal mouse mammary NMuMG and normal human mammary HBL-100 cells was used for comparison. RNA aliquots of 15 μg were separated in 1% formaldehyde agarose-gel electrophoresis and transferred to GeneScreen Plus™ hybridization membrane (New England Nuclear). Blots were hybridized with multiprime (Amersham) labeled inserts of either mouse (PM-4) (Saunders etal., J. Cell Biol. 706:1547- 1556 (1989)) or human syndecan (pUC19-hsyn4 Ba VW 1.1 kb fragment) (Mali etal., J. Biol. Chem. 265:6884-6889 (1990)), or with human growth hormone exons 4 and 5 (hGH) (Leppa etal., Proc. Natl. Acad. Sci. USA 59:932-936 (1992)) cDNAs, using the high stringency conditions suggested by the manufacturer of the membrane (New England Nuclear). All techniques based on modern molecular biology are fully explained in the literature such as in the laboratory manual entitled Current Protocols in Molecular Biology. Anchorage independent cell growth was measured in a soft agar colony assay. The six well-plates were first covered with an agar layer consisting of 2 ml DMEM, 0.5% agar and 4% DCC-FCS. The middle layer contained 10 4 cells in 0.5 ml DMEM supplemented with 0.33% agar and 4% DCC-FCS, with or without 10 nM testosterone. The uppermost layer, consisting of medium (2 ml), was added to prevent drying of the agarose gels. The plates were incubated at 37'C in 5% CO2 for 12 days after which cultures were evaluated and photographed. Tumorigenicity of S115 wild type cells, one hGH transfected control clone and two clones expressing human syndecan-1 was measured in nude mice. For this cells were cultured in DMEM containing 5% FCS and 10 nM testosterone. After four days in culture, cells were harvested with trypsin, washed, and 10 7 cells suspended in 0.2 ml of DMEM were injected subcutaneously into back of athymic male nude mice (balb-C). Silastic testosterone capsule, which is known to increase the growth rate of S115 cells (King et al., J. Steroid. Biochem. 7:869-873 (1976)) was simultaneously implanted. Nude mice were examined regularly for tumor development and the size of the palpable tumors measured at intervals. EXAMPLE II Growth factors enhance syndecan expression. NMuMG mouse mammary epithelial cells and 3T3 (NIH) mouse fibroblasts were routinely cultured in bicarbonate-buffered Dulbecco's modified Eagle's medium (DMEM; GIBCO) containing 10% FCS (GIBCO) and antibiotics, as previously described (Elenius etal., J. Biol, Chem. 265:17837-17843 (1990)). For experiments, cells were plated at equal density on culture dishes (Nunc) and grown to 60 - 70% confluency. Twenty-four hours before supplementing growth factor(s) to the medium, fresh medium containing 2% CMS-FCS (Vogel et al., Proc. Natl. Acad. Sci. USA 75:2810-2814 (1978)) was replaced. Equally treated cultures without growth factor addition served as negative controls. Porcine TGFB1 (R&D), recombinant human bFGF (Boehringer) and murine EGF (Sigma) were used in final concentrations of 2.5 ng/ml (100 pM), 10 ng/ml (570 pM) and 1.2 ng/ml (200 pM) respectively, in all experiments. For quantitation and isolation of cell surface syndecan, media were discarded at time points indicated in the text and the cell layers were washed twice with ice cold phosphate buffered saline (PBS). Cells were scraped with a rubber policeman into ice cold PBS supplemented with 0.5 mM EDTA and centrifuged. After subsequent washes by resuspension and centrifugation the cell numbers were measured by counting the nuclei with a Coulter Counter (Coulter Electronics). For quantitation of syndecan intercalated into the cell membrane, syndecan ectodomain was released by incubating washed cells in 20 μg/ml bovine pancreatic trypsin (Type III; Sigma) in PBS for 10 min on ice bath. After incubation the cells were centrifuged, leaving the ectodomain in the supernatant (Rapraeger era/., J. Biol. Chem. 260:11046-11052 (1985)). Sample volumes equal to 400,000 or 200,000 cells for 3T3 or NMuMG cells, respectively, were loaded on cationic nylon membrane (Zeta-Probe; BioRad) in a minifold-slot apparatus (Schleicher and Schuell), as previously described (Jalkanen etal., J. Cell Biol. 705:3087-3096 (1987)). Nonspecific binding was blocked by incubating the membrane for one hour at room temperature in PBS containing 10% FCS. Syndecan attached to the membrane was detected with a monoclonal antibody against mouse syndecan core protein (mAB 281-2) (Jalkanen etal., J. Cell Biol. 707:976-984 (1985)) that was radioiodinated by chloramine-T oxidation method (Stahli et al., Meth. Enzymol. 92:242-253 (1983)). The membrane was incubated overnight at 4 * C with 125 l-labeled 281- 2 in PBS + 10% FCS (10,000 CPM/ml). After five washes with PBS the bound antibody was visualized by autoradiography. The accumulation of syndecan ectodomain into the medium was estimated by taking samples corresponding to 1/50 (3T3 cells) or 1/100 (NMuMG cells) of the total volume of the remaining medium at selected time points. The samples were analyzed by loading them to nylon membrane as described above. The autoradiography signal was quantitated with a GelScan XL ultroscan densitometer (LKB) using GelScan XL 2400 software (LKB). For syndecan purification, cells were radiolabeled for 24 hours in low sulfate DMEM (MgCl2 substituted for MgSθ4; 2% CMS-FCS) with 100 μCi/ml 35 Sθ4 (New England Nuclear) in the presence or absence of growth factor(s). Cell surface trypsin-releasable material was collected, as described above, and after dialysis against Tris-buffered saline (TBS), the sample was loaded onto a 281-2-Sepharose CL-4B immunoaffinity column (Jalkanen etal., J. Cell Biol. 705:3087-3096 (1987)). Bound material was eluted with 50 mM triethylamine (TEA) (pH 11.5) and the amount of radioactive PG in each fraction was analyzed using cetylpyridiumchloride-impregnated Whatman 3MM filter discs (Rapraeger et al., J. Biol. Chem. 260:11046-11052 (1985)). For interaction experiments, fractions containing most of the labeled PG were pooled and dialyzed against PBS. To obtain unlabeled syndecan ectodomain for interaction assays (see below) the same procedure was used except that no radioactive sulfate was added to the culture medium and the syndecan containing fractions eluted from the immunoaffinity column were detected by immuno-dot assay using mAB 281- 2. The estimation of the molar concentration of syndecan was based on the use of previously determined syndecan concentration by total amino acid analysis (Jalkanen etal., J. Cell Biol. 106: ' 53-962 (1988)). SDS-PAGE and Western Blot - For western blot experiments cells were cultured 24 hours with or without growth factor (s). Syndecan ectodomain containing material released from the cell surface by trypsin treatment was fractionated on SDS-PAGE gradient (2 -15%) gel (O'Farrel, J. Biol. Chem. 250:4007-4021 (1975)). After electrophoresis, samples were transferred onto Zeta-Probe membrane using electroblotting 2005 Transphor apparatus (LKB). The syndecan antigen on the filter was detected with radioiodinated mAB 281-2 and the filter was washed, as described above for slot blot analysis. Northern Blot - RNA was isolated from 3T3 and NMuMG cells using 4 M guanidine isothiocyanate and CsCI density centrifugation (Chirgwin et al., Biochemistry 75:5294-5299 (1979)). RNA samples were size-separated on 1% agarose formaldehyde gel, transferred to GeneScreen Plus™ membrane (New England Nuclear) and hybridized with multi-prime (Amersham) labeled partial cDNA clone for mouse syndecan (PM-4) (Saunders et al., J. Cell Biol. 705:1547-1556 (1989)). After hybridization the membrane was washed in 2 x SSC and 1.0% SDS at 65"C (high stringency conditions). For rehybridization with glyceraldehyde-3-phosphate-dehydrogenase (GAPDH; Fort et al., Nucleic Acid Res. 73:1431-1442 (1985)) the bound PM-4 probe was removed as recommended by the manufacturer of the filter (NEN). EXAMPLE HI Induction of svndecan mRNA expression in the human breast cancer cells ■ MCF-7. growth-inhibited with toremifene. The steroid-responsive human breast cancer cell line MCF-7 was used to study the expression of human syndecan under different growth conditions regulated by estrogen and antiestrogen. For the experiment cells were plated at a density of 1.2 x 10^ cells/100 mm ø plastic culture dishes and grown as monolayer cultures in 10 ml per dish of phenol red-free DMEM medium with 5% dextran/charcoal stripped fetal calf serum (DS-FCS), 2 mM L-glutamine and 3 μg/ml insulin. For hormone-treatment 1 nM 17β-0-estradiol (E2). alone or with 1-6.25 μM toremifene, dissolved 70% in ethanol, were added to the culture medium on the day following plating. The ceils were cultured for 6 days, and the media were changed every second day. For RNA extraction the cells were washed in situ with PBS and scraped from the plates in 4 M guanidine isothiocyanate. EXAMPLE IV Treatment of Steroid-Responsive Tumors in Patients. Patients diagnosed as having a steroid-responsive tumor selected from a breast tumor, an endometrium tumor, a prostate gland tumor or a mesenchymal tissue tumor are administered a composition that contains efficacious amounts of the anti-steroid agent toremifene or tamoxifen, or efficacious amounts of the growth factor bFGF, TGF-β or bFGF together with TGF-β, in amounts ranging from 20-600 mg per day, depending upon the extent of the tumor, the patient's age, the patient's sex, and other treatments such as are taken into consideration in designing such chemotherapeutic protocols. The syndecan- inducing agent is administered for a period of time sufficient to induce syndecan in the tumor cells, such that the tumor cells now take upon a more differentiated phenotype and such that the growth of the tumor is arrested or significantly slowed by the treatment. EXAMPLE V Stimulation of Hair Growth in Epidermal Skin Cells. Patients diagnosed as being in need of increased hair growth in the scalp region are administered a composition that contains efficacious amounts _ of the anti-steroid agent toremifene or tamoxifen, or efficacious amounts of the growth factor bFGF, TGF-β or bFGF together with TGF-β, in amounts ranging from 20-600 mg per day, depending upon the extent of the needed hair growth, the patient's age, the patient's sex, and other treatments such as are taken into consideration in designing such protocols. The syndecan-inducing agent is administered for a period of time sufficient to induce syndecan in the epidermal cells, such that hair growth is significantly increased by the treatment. EXAMPLE VI Dermination of Mouse Svndecan Promoter and Enhancer Activities The mouse syndecan gene has been cloned and characterized up to -10 kbs upstream from the transcription start site. To determine the specific activities of different proximal promoter regions (up to -2 kbs from the start site) and enhancer regions (from -2 to -10 kbs) we have made plasmid constructs where these regions were cloned into pCAT basic or pCAT promoter vectors, containing the CAT reporter gene. The reporter CAT gene produces chloramphenicol acetyltransferase enzyme, which transfers the n-butyryl moiety of n-butyryl CoA to chloramphenicol. The n-butyryl chloramphenicol can be separated from native chloramphenicol by xylene. For the further characterization of the proximal syndecan promoter a series of retriction enzyme treatments was made on the upstream region (Hind III, Hind II, Bgl II, Stu I, Dra I, Cla I, BamHI and Pst I - Xho I) and the obtained fragments were cloned into the pCAT basic vector. For enhancer areas, three Xba I fragments were cloned into a pCAT promoter vector, where the SV 40 promoter was displaced by the Bgl II - Xho I fragment from the syndecan promoter. The plasmid constructs were transiently transfected into eukaryotic cells by calcium phosphate precipitation simultaneously with a β-Galactosidase expressing vector to determine the transfection efficiency. After a four hour incubation cells were treated with 15% glycerol and grown for approximately 48 h in cell culture medium. Cells were then scraped from dishes in TEN-buffer and the cytoplasmic extract was obtained by repeated freezing and thawing, β- Galactosidase acitivity was obtained in hte cytoplasmic extreact by adding 0.1 M sodium phosphate, 45 mM mercaptoethanol and 0.2 mg ONPG. This was incubated from 2 hours to overnight and the colour reaction was measured spectrofotometrically at 420 nm. The CAT activity was determined by adding 0.25 M Tris buffer, 25 ng n- butyryl CoA and 0.0626 μCi of 14 C-chloramphenicol to the cytoplasmic extract. These were incubated overnight, backextracted wiht xylene and radioactivity was measured by scintillation counting. The CAT activity was corrected for transfection efficiency by β-galactosidase activity. The cells used for proximal promoter constructs were 3T3 NIH, S115 (either hormone-treated of not) and nMuMG cells. For enhancer constructs we used 3T3 NIH cells grown in 2% CMS medium and to test the effect of growth factors in 2% CMS medium with 10 ng/ml FGF-2 and 2ng/ml TGFβ-1. All references cited herein are fully incorporated herein by reference. Having now fully described the invention, it will be understood by those with skill in the art that the scope may be performed within a wide and equivalent range of conditions, parameters and the like, without affecting the spirit or scope of the invention or any embodiment thereof. SEQUENCE LISTING GENERAL INFORMATION : ( i ) APPLICANT : NAME : Alanen-Kurki , Leena STREET : Piispankatu 6 E 31 CITY : TURKU COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-20500 NAME: Auvinen, Petri STREET: Kirschgartenstrasse 5 CITY: HEIDELBERG COUNTRY: GERMANY POSTAL CODE (ZIP): D-69126 NAME: Jaakkola, Panu STREET: Kellonsoittajankatu 13 B 20 CITY: TURKU COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-20500 NAME: Jalkanen, Markku STREET: Rauvolantie CITY: PIISPANRISTI COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-20760 NAME: Leppa, Sirpa STREET: Valkintie 14 as 6 CITY: LITTOINEN COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-20660 NAME: Mali, Markku STREET: Inkereentie 176 CITY: SALO COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-24280 NAME: Vihinen, Tapani STREET: Kaskenkatu 11 C 54 CITY: TURKU COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-20700 NAME: arri, Anni STREET: Juhaninkuja 2 as 2 CITY: LIETO COUNTRY: FINLAND POSTAL CODE (ZIP): FIN-21420 (ii) TITLE OF INVENTION: SYNDECAN STIMULATION OF CELLULAR DIFFERENTIATION (iii) NUMBER OF SEQUENCES: 3 (iv) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Floppy disk (B) COMPUTER: IBM PC compatible (C) OPERATING SYSTEM: PC-DOS/MS-DOS (D) SOFTWARE: Patentln Release #1.0, Version #1.25 (EPO) (v) CURRENT APPLICATION DATA: APPLICATION NUMBER: WO TO BE ASSIGNED (vi) PRIOR APPLICATION DATA: (A) APPLICATION NUMBER: US 07/988427 (B) FILING DATE: 01-DEC-1992 (2) INFORMATION FOR SEQ ID NO: 1: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26700 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: both (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iii) ANTI-SENSE: NO (ix) FEATURE: (A) NAME/KEY: CDS (B) LOCATION: join(4378..4443, 22026..22106, 23001..23483, 23905..24039, 24251..24418) (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 1: TCTAGATATT CAAACTCACC AGATGGAGTG ATGTCCACCC CTATTGGTGG GAGTGACTAG TCTTTCCTCT GTCTTCTGAC TCAGATGCTT AGCTAGCTCT TTAGGACCCA CCCTCACACC TGCAAATAAT ACTTTATTTG CTCTCTTAGT ACCTTTAACC CAGTGGAGTT GACATGAGAA ATTAACTACC ATAATTTATA ATATTTCATT TCATAAATGA AAAGTAAAAT AAATTAAAAA ATAGAAAGGT TTGAGCATGA TGGCCCAGTG GTAAAGGCCA GTGGCTCCAA CGCAAGTCCT GACAAATGGT AACGGGCCTG TTCTTCAGGC TTGAGGGAAG TTTATTGATT GAGGCTAAAA GCAACCCAAA GGCTCCACTT GCCTAGTGTG AAGCCCTGGA TGTGCTCTCC CACACTGCAT GTCCACCTGT GGTGTCAGCA CCTGGGAAGC TGAGGATGAT GGGGAGTCCA AGGTCATTAG CTACATAGTA TAGGCTAGCT GGGGTACATG GGTCACAAAA AAGAAAAAAA AATAAGCACA TTGTAATCCC AGCACTTGAC AGACCAATGG GGGGGGGATT GCTGTGAGTT TAAGACAGCC TGGCCTACAA AGAAAAACCC TACCCAAACC CAAGAAAAAT GAAACCAGTA ATATAAATAG CTATTTTCAT TTTAAATGCT CTAAAGACAC AGCGTTAACA CAAAAGCTCT CGTCTGTGGT TCCTATTCCC TCCTTCTCCC CCAGGTCTTC TTTAATGTAT ACTTTTTGTT TGCTTATTTG CTTGTTTTGG ATTTTGGCTT TTAAAGACAG GGTCTCACTA TGTAGCTCCA ACTATTTGGG AACTCACTAT GTAGACCAGG CTAGCCAGGG ACTTATAGAG ATCTACCTAC CACTGCCTCC CAAGTGCTGA GACTAAAGGC ATGTGACACT TTGCTTGGTT ATTACAAACA TTTTAAAAGA ACATTTTGAA CATTAATAGA TGTATGTATA TATATCACTC TATGTAGTAT ATATGTTAGA CATTTTTCAC TTGAGATACA TATTTACTCT CAAAATAAGT TTTTTGTTTT TTTTTCTTCT TTTTAAATTT ATTTTATTTT TTTTTTATTT ATTTTATTAT TATATGTAAG TACACTGTAG CTGTCTTCAG ACANACCAGA AGAGGGAGTC AGATCTTGTT ACGGATGGTT GTGAGCACCA TGTGGTTGCT GGGATTCGAA CTCTGGACCT TCCGAAGAGC AGTCGGGTGC TCTTACCCAC TGAGCCATCT CACCAGCCCC TTAAATTTAT TTTTATCTTA TGTCCATTGG TGTTTTGCCT GCATGTATGT GTAAAAGTGT CAGAAACTGA AGTTACAGAC TGTTGTGAGC TACCATTGTT GTGGGTGCTG GGACTTGAAC CTGGGTCCTC TGGAAGAGCA GTCATTATTC TTAACCACTG AGCCATCTCT CTAGCCCTCG TTTTTTAGTT TTTTTTTTTG TTTTGTTTTG TTTTTTGTTT TTTTAAGATT TTCTTATTTA TTATATGTAA GTACACTGTA GCTGTCTTCA GACACTCCAG AAGAGGGCGC CAGATCTCGT TATGGATGGT TGTGAGCACC ATGTGGTTGC TGGGAATTGA ACTCCAGACC TTTGGAAGAG CAGTCAGTGC TCTTAACTGC TGAGCCATCT CTCCAGCCCC GTTTTTTAGG TTTTTGAAGA CAGGGTTTCC TGTGTAGCTC TAGCTGTCCA GGAACTAGCT CTGTAGACCA GGTTGGCCTC AAATTTAGAG ATTTGCCTGT CTCTCTGCCT CTCGAGAGCT GGGATTAAAA GTGTGCAGCC CAACAATCTA CTCAAAGTAG GTTTTGAAAA AGCTTTCCAT ATTAGGAGTT AACTAGCTTC ATTTCAGAAA TACTGCATGG AATTCAAATG TGGGACCATT CATAGCTACT TTGGTTTTCC TTCAGTGACA GGCATTCGGC ATGCCTATTA GGGAAGTCAA ATGGCCTGGA GAAGTCATCC TGGGTGAGAG GGCTAATGCA TTTTCAGCTT GACAGACACT GTCAACCTAT GCAGACAGTC TGCTCCAGCT CAGATGTCAA TTGCATGCAG ACCTGCAGTC AGACGCTAAG CTCCCTACCT ACTCTCCATC AGCTTAGATG TAAGGGGTGC TGGAACAAAG GCTCTCTCTC TCTCTCTCTC TCTCTCTCTC TCTCTCTCTT TCTTAGAATT AGTATTCTAT TTTATTTTAT GTAAATTGGT ACTTCACTTA CATGTATGTC CGTGTGAGGA TGTTGTATCC TCTGGTACTG GAGTTATAGA CAGCTGTAAG TCGCCATACA GGTGCTGGGA ATTGAACCCT GATCCTCTGG AAGAATAGTC AGTGCTCTTA ACCCCTGAGC CATCTCTCCA ACCTCTTGCA TATTGAGGAC AGGGAGGAAT CACAAGCCAT GTAGGGTGCC TGGGCTCTGA GGTCAACAGG ACCATAGCCT CCTTTCTTTA TGTGCCTTTC TTGGGGTCTC CCTATAGGAG TCGTCTTCGT TGCCTCTTTA CTGTCTCATT GATCTGGGCT AAACTTATGC AGTTGGAAGG AAAGATCAAG CTGGTCATGT TTAAAACATG AAACAGCCTC ATCAGTTCCC TTCCTGTTCC CGTCTCCCCC CCCCCTCCCG CCCCCATTTT GAGAGGACAG GAAGGTAAAA TACCAAAGTG TCCTATTTTC CTCCAAATAT CAGGCTCAAA GGACTGAAGA GCTGACTTCA GATCCCAAAG CCACTGTGTT AGGAGGCACC TGCTTTTTAG GTCCTAAGCC TTCCTGAGCC TTGCTATTGG GTATTCTTTA CCAAGACCCT CAAGGATCTA GGCAAGAACT GGGCAGGATC TGTATGTAGC CCATAGTTAG ACCTAGGGCA GCTGAGACGC CAAAAGGGAG AGTTTCCTGA GGACAAAAGT GTTCAAACAC AACTGGGTGC TGGTTGTTGG GCTACTCGTG GAGGTGTGGT GTGTGTAAAG GAGGCTGTTG AATTCCCAGA AGGCTGGTTC CACAGTGTAG AGTCTACACT GGGGACTTCC CGAGACGCTG AGCCTCAGAT CTAGCTTCTC AGTCCAGGCC AGCTGATGTG GGGCTGAGGA ACAAGGATGG ATGCCATCTA TGGCCCTGCC TTGCAGGTGC AAAGGGCCTT TGGCACCATC TACAGATTGA GGGCAAGACA GGGCTGGTTC TTCCTCCTTG CTCTCGCTGC TATCTGCCTC GCCTGTAGGC TCTCTGGGCT CCTTTTTGGA CTGACACGTC TGAAGGAGCT TGGAAACTGT GAGGTCCAGG CCCCATAGAG AATCATGAAG GAACAGGAAT TCAACTGGAG CTCCGCAGCT GGTTAGGCCT GCGGTCACCT GGAAACAAAG AGGCCATTTA TTTTTTCCTT TGGTCTTGGA CAAGGAAGAG AAGGGGCTTT CTATAAATAG AAAGACAGCA AAAAAGAAAA TAATAATAAT AATAATAATA ATAATAATAA TAATAAAAAC AATAACAAAG CCAGCTCTTC CAGACAGTGC TCATGTCTTT AAAGGTCTTT AAAGGTCTGG AGTTCCCAGC AATTAAGTAA AGGACCAAGA CCTCAGGGGT CCCCTATCCT CAGCCCGTGG GGAGGTGGGA ACCATACATC GATCCCTCGG TTTATATATA GCCTCATCGC TGTGGGGCTC CGAGGTTGCC CCCAAAATCT TGCTCACCTG GAGGACCCCT GGGTGTCCTC GCCCAGAGGG CGCTGCAGCC TCGCACGTAG AGAACTAACA TCGCCCTTCT CCAGGGCAGT GCCTCCGGAC TCCGGACCAG GACATAGTAG CGAGTGCACC TGGGTCTCCG TCAGCTACGC ATCAAGGAAG GTGCGACGCG GGAATTACAG ATTGCCGGCA CTCACCAGTG CTCAGGGGAG GAAGGTGGGA CTCAGACCTG CAAGAGCTGA AGAGTGGGGT GGGCTTCGAT CCTAGGAGGC GTGGAAGGGG GTGTGGCTGG ATCCCTGGGG GGTGGGGCGC TCCAAGGGGC GGGGCAACCC AGGGGGCGGG GCCCGAGGGG TGGAGATTGG GACTACCCAG GCCCGCGGAG CTGGGGGTGG GCGGCTAGTT TTGCAACTGC AGAGCCTTTG GGTTTATTAT AAGGCGGAGC 4 TCCGCGGGAG AGGTGCGGGC CAGAGGAGAC AGAGCCTAAC GCAGAGGAAG GGACCTGGCA 4 GTCGGGAGCT GACTCCAGCC GGCGAAACCT ACAGCCCTCG CTCGAGAGAG CAGCGAGCTG 4 GGCAGGAGCC TGGGACAGCA AAGCGCAGAG CAATCAGCAG AGCCGGCCCG GAGCTCCGTG 4 CAACCGGCAA CTCGGATCCA CGAAGCCCAC CGAGCTCCCG CCGCCGGTCT GGGCAGC 4 ATG AGA CGC GCG GCG CTC TGG CTC TGG CTC TGC GCG CTG GCG CTG CGC 4 Met Arg Arg Ala Ala Leu Trp Leu Trp Leu Cys Ala Leu Ala Leu Arg 1 5 10 15 CTG CAG CCT GCC CTC CCG GTGAGTGTGG CCCGGGGCAG GGCTGGGAGG 4 Leu Gin Pro Ala Leu Pro 20 CGGCGGGAAG CCGGGACTCG CCACTCGCCG ATGCCATGCA GGCGGCAGCA CGTGGAGGGG 4 GAGGGGAGCG GGGACTTCTT CCCGCGCTGC CTGGCGGATC CTGGGATGGT GAGCCCTTTA 4 ATGAGGACTC CTGTCCCAAT TCCTCTACGG TCCGTGGATG CCAGGAGGCT ATCCCAGCTC 4 GTGGTCCGGG CGTCCTGCAG AGTGGAACCT CCATTGGTTC CCCGCTCCCA ATTAAGTAAA 4 ACGACTCCAC AGGGGTCTGA GTCGCCGGCC TTAGGCGCTC CGCCGGCCTT AGGCGCCGCT 4 TGGAGTTGCT CTCTCCCGTT GCTGTCTTGC TGGCCATCTC AGCGGCCTGG CCTCCGCCAG 4 TGTCCCGGAG GATGCAGTGG CCATGGCCAA ACGCCTTTTC CATAGACCCT AATTCAAACC 4 AGACTGCAGG CTGCACCCCC AGCGCCGCGG AGTCCGGGCG CTCGGCCCTT TGCACCGGGG 4 CAAGTTTGGG CACAGCAGAG CCGGCGCGGG AACAGGGGGA AGCTGACGTT CGGGGTGGCG 5 GGAGGGACGG GATTAAGGCT GTTTGTGGGA CACAAGAGGG TGGCTCAGGG ACTTCGGTTT 5 TTCTCTGGCT GCCCCAGGTG AGCCGGGCCG AGCTGGCAGC GGGAGGTTCC GGGAAGTTGG 5 CTTCAGAACG CTGAAGACCC TAAGAACCCA ACTTTGGGGT CGCTGAAGTT GTGCTGCCCC 5 CGGAGGGCCT CCTCCGCATG GCCCGCGCGG GGGACCCTCC CCGCGAGTGG ACCCCGGTAC GGCTCTTCCC CTCCCCCGAC TCGGCTTTGT GCTGAAGCCG CGCGTAGGGA AGGCGGGTCC 5 CTTGGCCCGC CCAGTAGGGC CGCGGGGAAA GAGGGACGAA CGTGGAGCTG GCGACTGGTG 5 GGGGAAGCTT CTGGGTAGGA TGCAGCCATC CACCTTTGGT GGGGTCGGTC TCTCTAATCA 5 GCGGCTTGGC GACAAAGAGC TTGGTCGAGG GTACCCCAGA AAGTGCTCTC CCGCCCCAAG CCGCCGTCGC TAGCCCGCCT TCCCAACGGG CGCTTTGTTC TCGGCCCCTG TAACCCTTCC CTGGGAACCG CCCCGCAGCG CTGGTCCTTG ACGTGGGCCG GGTCCTGGGT CGCCGCCAGT GTCAGCGCTG CCCTCCGGTG TCCACGCCCC TAGCCCCCGC ACCCGCTGTG AAGTCCCGGG TGTCCTTTCC ACTGGCGCTT TGCCCAACCC CTGGAAGGCA GAGGCGAGGT GCGGAGCCTC AGGCTTTATC CTCCCGGAAG TGGCAGTCTC CCACCGCCAC ATCTGGTCTG CTTAACTTCG ATAGTCCTGG CAAAGGCAGA CACGTGCACA GGGAAGGAGA GTTGAGCGCT GGTAGATACC AAGGTCGTGT ACAAATAAAG TGGCACACGA CACGTCCCCA GTCACTGTTA ATGCATTGCC TTCGCTCCTT CCCAGGTGGC TGGTGCTCTC CATCACTCTG GAGCCCAAGA GAGGGCCTCC ATAATTGTAT TGCCCATGAG TTGGGGTTGT GTGGGGGCGC CAAATCAGGG TTCTCTGGGA GGGCTATGAA TTCCGAACTG AGTCTCCTGT GCACTCCTGG CTTTAAGGTT CAAGAAATTG TTTGAGGGTT GTGGTTTTTG TGGGACTCAG ATTATGCCTG GAATCATAGT TACCACTGTG GAGAAGAAAG TGGAGC ACT TAGCATGCCT CCCCGGCCCG CCTGGCATTA CCTCCGGCTC TGTTCTCTAG GCCCAACGTG AGGCCTCACT GGGGCAGTAC AGATGCAGTA CTGAATTTCT TTCCAGCCAG GATCTGGAGA GGTGGTGTTC TCTTCCCTGG TGTCTTTAGA GAGGCAGATA TTCCTGTGAC CTAAGCCCCT CAAGCACCCA TTAATAATGC TGAGTAGACA ACTAGAGGTG GCGTTTTCCG GAACTTCCTG TGTGCTGGCC TGGGAGGTTG AACCCTCTAG GAAACAGGTC TAGGAAGTAG AATTATCTCA ATGGAAGGCT TCCTGGAGGA AGAAGATGAG CTGAGCCCCC AGGTCACTGT CTGAGCTTTA GGATCAGACT CCCACTTGGA GGCAAGAGTG TTCGTTTTAC TTTTTTTTTT TAAGTTTAGT TTATTTTCTC TCTAACAGAA AACAAACAAA CAAACAAAAA AAAACCCCAC ATTGTTTAAA AGTGGGTGCA TAAGAGTGAG GACATATTCA GAGCTTCCCC TTTTCCTGAA AAATGAAGGC AGCTGGGATT TACTTAAAAT GAGAGCACAT ATCACAATTG CCAGAGAGCT GGTCCCTTTC TCAGGGCTCC CTAAGCTCCT GTGGGAAGCA GGTCAGACAG CCCTGGGGAC CAGAGAGATA GGGAGTGCTT TTGGGTGCCT GCCTTTGAAT GGGGAAGGGG GGGGGAGCTG CTGGGATCAG AGGCTGCTAG CAACTACTCC CCAGAGACTG AAGCAGGTTT GTCCCTCAGT GTCCTGTGGT CTTCTGTTTC TCCTATATAG AATAGGAGAA ATGGTTATTT GCTCTGGAAT AGTGACTTGC TATTTGTTCC CTTTCTTTCC TCTCCCTTAC TGTAATCATT TGGACTAGTA GAGACACTTT CCCCAGGTCT GGCAGAATGG GAGGGAGTGG GGGAGGCCTG TGCTTGCATG ATGTCACTGC TGGCTTCAGC TCTCCAGGGA GGGTGGAGTT GGTTGTAACC TACCTGTGGC TCTTGATGGG CCACAATAAA ACCTCATTAA CACACATTGG TAGGGAGAAG GGACTGGAAA GAATGATGGG AAAGATTGAT GTTTTTCCTT TTTTTTTTTT TTTTTTTTTG GCAGTACTTT CTAGATCTCC CCTCCCCCTT GCTGCAGCAA AATTTTGGAT TCCTGAAGTC CTTTGAGAAT GTATAATGGT AGCCAGACTT TTTTTTTTTC AGTCAGCTCA AAATTGCCTC CTTATAAAGT ATCCTTGGTT GTTTTTTGTT GTTGTTGTTG TTGTTGTTTT GTTTTGTTTT AAGACAAGGT TTCTCTGTAT AGTCCTGGCT GCCCTGGAAC TCAATATGTA GACCAGGCTG GCCTCAAACT CAAAGAAATC CACCTACTTC TAACTTTCAG TGCTGGGCCT AAAGGTGTAG GCCACCAAAA GTGCTCAACT TTTACAAAGC AGTCTTACTT TGAGCAGGAT TCTGAAACCC TTATTTCCTT TCTGTTATCT TCAACAATAC ACTGCTAGGT GTATTTAGTC CCTCATGATG CTGGGCCTCC TCAAGTGGCG CCAGGTCAAG CAGTCTCCTG GTTTTTGGTG GCTCTGAAGA AGACTGTGTC CCAGTGACTG GCAGTTTGAA TTCGGAGCTT CTCTTTTCCT TCTCAGTCTT TGGCAGGCAG AGTGACACTG GTGTGCCCAA GCCTGGAGCT TCTCTGTTTA ATTCTAGTTT ATTTTCTTTA TCAGACTGAA AAACAAATCA GGTTGGTTAT AATTCTTATA AACACGAAGG TCTCACCTTT GCGTACGTCT CCGGCTGTGT GGGTCTGATG TCCCTCGGGA ATCTCTGTTG AGGCTGCTGC AGTGTGTGTG CGTGTAGAAA GGGCAAGGTA GAATGGACAG AAGCGTGCTG CCCACCCCAC TGTCCTGTTC CTAAATGATG AAGCACTGGC CCGGTGAAGA GCCTAGAGAA CTCCCTCGGT GGGAGATGCA CACAATGCCA GGAAGCACAC AGGAGCTTGA GTTCCAGCTT GGCAGTGTCT TCTCTTTGGT GACTTTATCA GCTCCAGCTG CCCTGGACTA ACAAACAAGG CTAGCTCACT CTCAGTATTG ATAATCGAAG GTCCTTGGTT CTGTTTGAGA CTGATCCTCA CTCGGTAGCC TTGAACTCTT AGCAATTCTC CTGTCTCAAC TTTCAAAGAG CTGAAATTAC AGACTCGAGC CACCATATGC GACTGAAACC TTGTTCCTAA TCCTTGACTG TGAACGACTC TTGGGTTTGG TTCTTTCTCC ATTTCTTTAG TGTATGTTTT AGTTCGCGTC CTACATAATC TATTGCCCAT ACTTAGAAAC AACAGGTTAG AGACAGCATT GGGTCCAGCA GAGCCTCACA CTGAAGCTCA GTCCTGCCAC TGATTTACCG TGTCAGCTCA AGTGACTCAC TTCCAACTCC TCTGCTCCCC ATCTGTAGAG TAGACATCAC CATACCTGCT CTTTCTGCCC ACATTCTGTC ATTAACATGT TCATTTCATA ACGATGGTGC AAAAGTGCTT TGTAAGTAAA GTGCTGGGGA AATGTTAGCT GTCGATAATG GTTAGGGTTA ACTTTTTATT GAGTGCCTGT TGTGTGTGGG GTTGGGTGGG GTTTTTTTAG AGGCTTGGTA GTTTTCTTAC TTCTTTCCTA CTTAGCTTTT CTTCCTAAGC CTTTATGGTA TGTATCATTG CCTGATTGTT TGAGTGTGTG CACTGAGGCA CGCCTGTGCA TGTTTGAGAG TATGCTTGTG CGTGCTCTCG TGCTCACATA TGTATGGTGT GAATACACTG TAGAGTGCAG GCCGGCACAC TGGGGCTGGC TGAATCCTGT GAGCCCTGCC TGGAGTTTGC AGATCTTCCT TGGACACTCC TGCTTGTGAG CATTTTGTGT GGAGTGACTG TTTAGCTGGC TGTAGCC AC ATTGTGCCTT TGGGTAAACC CTGAGTATTG GGAAACACCC GCTGCTGTTG CCAGGGAGAG CCCATCCCAT TCACTTCAGT CTCCTTAATG TAGAAGCCTT 2 GTCGAATTAG CTTCCACTGT CCCCAACCCA AGAGTACCCT GTCCTTTCTT CACTAAGAAG 2 GCCAGGATAC AGTCCTTCCT GTGGCTGATA AGACAGGCCT TGGGACAAGG CCTGGGACCA 2 CACTGTGTGG GCAAAGCTGC TTCAGCACCG ATGGCTCCTC CATGCCAAGC TTGGCTCTGC 2 TTCTCACAGT TGAGACTTCT GTGCGCACAC CCACTGTCTA GCTCAGCTGG ACACTGATTT 2 TCTTTAAATG TATAGATTTT GGGGTGGGGT GTGCTGAAAG CTCCCACTGA TGCCCCAAGC 2 CTGAGTCTCA GAGTATGATC AATTGATGGC TTTCATGGGT ATCACAGCTT CTGTTCCCAG 2 GTCAGACTCC CTGACCAGTC AGAGCATCCT GGGGTTAGAC AATGTCCCCG TCACTTGTGC 2 CTCCACCTGG CACCAGGCTA TGATGTTATG GCATTGAGGG TATGAGAAGG ACCAGGGGTT 2 TCCCAGAGTT ACGCCCAGGC GCACAGGCAA TTGTTTCCTA CATGTGTGGC TGGAATGGTT 2 GGGTGAGCCT TTTCAGCTGC CTACAATAGG AACCCAGGGA TACTGGGCAT TGACCAAGGC 2 ATATCTCATA CCCTTTTCTT ATCTTTCTGC AG CAA ATT GTG GCT GTA AAT GTT 2 Gin He Val Ala Val Asn Val 25 CCT CCT GAA GAT CAG GAT GGC TCT GGG GAT GAC TCT GAC AAC TTC TCT 2 Pro Pro Glu Asp Gin Asp Gly Ser Gly Asp Asp Ser Asp Asn Phe Ser 30 35 40 45 GGC TCT GGC ACA GGTAAGACTG ACCCAGAACA CTGAGATGGC ATAGATCATG 2 Gly Ser Gly Thr TGAGTTCAGC CTTTGCTAGG TCACCTTTGG GGTCTCAGAA GGCTTCAGCT CCTGGTAGAG 2 CATGAATCAC GTCAGGCGTG ATGCTGGAGA CCTCTCCTAC CCTGACACCC CAAACCCCCA 2 CCTCTGACCC TGCA GGT GCT TTG CCA GAT ACT TTG TCA CGG CAG ACA CCT 2 Gly Ala Leu Pro Asp Thr Leu Ser Arg Gin Thr Pro 50 55 60 TCC ACT TGG AAG GAC GTG TGG CTG TTG ACA GCC ACG CCC ACA GCT CCA 2 Ser Thr Trp Lys Asp Val Trp Leu Leu Thr Ala Thr Pro Thr Ala Pro 65 70 75 GAG CCC ACC AGC AGC AAC ACC GAG ACT GCT TTT ACC TCT GTC CTG CCA 2 Glu Pro Thr Ser Ser Asn Thr Glu Thr Ala Phe Thr Ser Val Leu Pro 80 85 90 GCC GGA GAG AAG CCC GAG GAG GGA GAG CCT GTG CTC CAT GTA GAA GCA 2 Ala Gly Glu Lys Pro Glu Glu Gly Glu Pro Val Leu His Val Glu Ala 95 100 105 GAG CCT GGC TTC ACT GCT CGG GAC AAG GAA AAG GAG GTC ACC ACC AGG 2 Glu Pro Gly Phe Thr Ala Arg Asp Lys Glu Lys Glu Val Thr Thr Arg 110 115 120 125 CCC AGG GAG ACC GTG CAG CTC CCC ATC ACC CAA CGG GCC TCA ACA GTC 2 Pro Arg Glu Thr Val Gin Leu Pro He Thr Gin Arg Ala Ser Thr Val 130 135 140 AGA GTC ACC ACA GCC CAG GCA GCT GTC ACA TCT CAT CCG CAC GGG GGC 2 Arg Val Thr Thr Ala Gin Ala Ala Val Thr Ser His Pro His Gly Gly 145 150 155 ATG CAA CCT GGC CTC CAT GAG ACC TCG GCT CCC ACA GCA CCT GGT CAA 2 Met Gin Pro Gly Leu His Glu Thr Ser Ala Pro Thr Ala Pro Gly Gin 160 165 170 CCT GAC CAT CAG CCT CCA CGT GTG GAG GGT GGC GGC ACT TCT GTC ATC 2 Pro Asp His Gin Pro Pro Arg Val Glu Gly Gly Gly Thr Ser Val He 175 180 185 AAA GAG GTT GTC GAG GAT GGA ACT GCC AAT CAG CTT CCC GCA GGA GAG 2 Lys Glu Val Val Glu Asp Gly Thr Ala Asn Gin Leu Pro Ala Gly Glu 190 195 200 205 GGC TCT GGA GAA CAA GTGAGTGGCT TTGCATTTCC TGGGTGGCCA CTAGTGCCTG 2 Gly Ser Gly Glu Gin 210 CACCTGGCCG CCTAATGTCC TCATTACAGT GACAGGTGAC AGGGTCCCAC CTTCCTCCTG CCCGAAACAG ACTGATTGCA AGATCAGGAG GTGGGCGACT CCTTAGATGT CATTCAGGAG CTTACAGCAG GGTGAATTTT CCGTCTTAGA CCTTCATGGG AATTTTCACA CAACAATGTG TACGTTGTGT CACTGGAGGC GGTATCTGTG TCTTGGCCTG CCAGGGTCCC AGGTGTGACT GACTGCATTC CTTGACAGAT GCTGGTATAG GTTGGCTACG TCTGATGGGG GTGGCAGGGG ATCCCATCAG GTATGGCACT GCTCAGGTTG CTGTTGTGTC AGTGGCTCCA GCTGACCTGA 2 TCCCAACCTA CCCTTCTGTA G GAC TTC ACC TTT GAA ACA TCT GGG GAG AAC 2 Asp Phe Thr Phe Glu Thr Ser Gly Glu Asn 215 220 ACA GCT GTG GCT GCC GTA GAG CCC GGC CTG CGG AAT CAG CCC CCG GTG 2 Thr Ala Val Ala Ala Val Glu Pro Gly Leu Arg Asn Gin Pro Pro Val 225 230 235 GAC GAA GGA GCC ACA GGT GCT TCT CAG AGC CTT TTG GAC AGG AAG GAA 2 Asp Glu Gly Ala Thr Gly Ala Ser Gin Ser Leu Leu Asp Arg Lys Glu 240 245 250 GTG CTG GGA GGTGAGTCTT CTTTCAGGTG GAGAGGAGGA GGCAGGTGGT 2 Val Leu Gly 255 GGCTCTGAGG TAGCCTGGGT TGCTGGGGTG AAGCATCTTT AGCAGCAGGG TGGGGAAGGA 2 GGAGGGTCAA TTCTACTCCA GGCCACCTCC TAGGCTGTCC GTCTAGTCTG GGAGAGACTA 2 CCACTGACCC CGTGGAGCTA CTGATCTGAG CCTGCCTCTG TTCACTCCCT A GGT GTC 2 Gly Val ATT GCC GGA GGC CTA GTG GGC CTC ATC TTT GCT GTG TGC CTG GTG GCT 2 He Ala Gly Gly Leu Val Gly Leu He Phe Ala Val Cys Leu Val Ala 260 265 270 TTC ATG CTG TAC CGG ATG AAG AAG AAG GAC GAA GGC AGC TAC TCC TTG 2 Phe Met Leu Tyr Arg Met Lys Lys Lys Asp Glu Gly Ser Tyr Ser Leu 275 280 285 GAG GAG CCC AAA CAA GCC AAT GGC GGT GCC TAC CAG AAA CCC ACC AAG 2 Glu Glu Pro Lys Gin Ala Asn Gly Gly Ala Tyr Gin Lys Pro Thr Lys 290 295 300 305 CAG GAG GAG TTC TAC GCC TGATGGGGAA ATAGTTCTTT CTCCCCCCAC 2 Gin Glu Glu Phe Tyr Ala 310 AGCCCCTGCC ACTCACTAGG CTCCCACTTG CCTCTTCTGT GAAAAACTTC AAGCCCTGGC 2 CTCCCCACCA CTGGGTCATG TCCTCTGCAC. CCAGGCCCTT CCAGCTGTTC CTGCCCGAGC 2 GGTCCCAGGG TGTGCTGGGA ACTGATTCCC CTCCTTTGAC TTCTGCCTAG AAGCTTGGGT 2 GCAAAGGGTT TCTTGCATCT GATCTTTCTA CCACAACCAC ACCTGTTGTC CACTCTTCTG 2 ACTTGGTTTC TCCAAATGGG AGGAGACCCA GCTCTGGACA GAAAGGGGAC CCGACTCTTT 2 GGACCTAGAT GGCCTATTGC GGCTGGAGGA TCCTGAGGAC AGGAGAGGGG CTTCGGCTGA 2 CCAGCCATAG CACTTACCCA TAGAGACCGC TAGGTTGGCC GTGCTGTGGT GGGGGATGGA 2 GGCCTGAGCT CCTTGGAATC CACTTTTCAT TGTGGGGAGG TCTACTTTAG ACAACTTGGT 2 CAGGATTACA AGTATTGCTT GCACATTGAG GG 26 (2) INFORMATION FOR SEQ ID NO: 2: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 311 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 2: Met Arg Arg Ala Ala Leu Trp Leu Trp Leu Cys Ala Leu Ala Leu Arg 1 5 10 15 Leu Gin Pro Ala Leu Pro Gin He Val Ala Val Asn Val Pro Pro Glu 20 25 30 Asp Gin Asp Gly Ser Gly Asp Asp Ser Asp Asn Phe Ser Gly Ser Gly 35 40 45 Thr Gly Ala Leu Pro Asp Thr Leu Ser Arg Gin Thr Pro Ser Thr Trp 50 55 60 Lys Asp Val Trp Leu Leu Thr Ala Thr Pro Thr Ala Pro Glu Pro Thr 65 70 75 80 Ser Ser Asn Thr Glu Thr Ala Phe Thr Ser Val Leu Pro Ala Gly Glu 85 90 95 Lys Pro Glu Glu Gly Glu Pro Val Leu His Val Glu Ala Glu Pro Gly 100 105 110 Phe Thr Ala Arg Asp Lys Glu Lys Glu Val Thr Thr Arg Pro Arg Glu 115 120 125 Thr Val Gin Leu Pro He Thr Gin Arg Ala Ser Thr Val Arg Val Thr 130 135 140 Thr Ala Gin Ala Ala Val Thr Ser His Pro His Gly Gly Met Gin Pro 145 150 155 160 Gly Leu His Glu Thr Ser Ala Pro Thr Ala Pro Gly Gin Pro Asp His 165 170 175 Gin Pro Pro Arg Val Glu Gly Gly Gly Thr Ser Val He Lys Glu Val 180 185 190 Val Glu Asp Gly Thr Ala Asn Gin Leu Pro Ala Gly Glu Gly Ser Gly 195 200 205 Glu Gin Asp Phe Thr Phe Glu Thr Ser Gly Glu Asn Thr Ala Val Ala 210 215 220 Ala Val Glu Pro Gly Leu Arg Asn Gin Pro Pro Val Asp Glu Gly Ala 225 230 235 240 Thr Gly Ala Ser Gin Ser Leu Leu Asp Arg Lys Glu Val Leu Gly Gly 245 250 255 Val He Ala Gly Gly Leu Val Gly Leu He Phe Ala Val Cys Leu Val 260 265 270 Ala Phe Met Leu Tyr Arg Met Lys Lys Lys Asp Glu Gly Ser Tyr Ser 275 280 285 Leu Glu Glu Pro Lys Gin Ala Asn Gly Gly Ala Tyr Gin Lys Pro Thr 290 295 300 Lys Gin Glu Glu Phe Tyr Ala 305 310 (2) INFORMATION FOR SEQ ID NO: 3: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2196 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: both (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (genomic) (iii) HYPOTHETICAL: NO (iii) ANTI-SENSE: NO (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3: TCTAGAACAC TTATTAAGAG CCAGGCACTG AAAAGTGCAG ACTCCCTCAT TTCATCCTGG CCGTGCTTAC AAGTAGTTTC CATGCTCTGG TAACCCTGTG CAGAGGGCAG CGTGGGAGGC GGGCCGCTTG GTGGACGGTC ATGGGGGCTC TGCATGGGTG GTTGCCCTTG CCTCAGAAGA ACTCCCTAAG TAAGAGCAAG TTAGCCTCCC TAACCCCTGG TGGGTTGTTG CTTCTTTTCT CCTCTTGTTT CTGCCAAGAG AGGGTGGACC AAGAAGACCC CAGCCTACAG AACATGTGAT CCAAATAAAC TTCTTTTTAG TATAAATGTC CTAGCCTGTG ACGTTCTGGT AGACTAGCAC AAGATGGACC AAGACAACTC TCATCGAGAC TCTGAGGAAC GAACTGGCAT CACATGGGAA CAGGAAATGA AGCTTAGAGA GAGGTTCTGT GGCTTGTCCA ACATGGCTGT AGTTTAAATC CAGCTTGCCA CCAAAGCACA CACATTTCAC TGCTGTGCTG GGCCGGGCCT CAGATCCCAG GGGCTCCGGA GCTAGAAGGA CACGTGTATC AGCCATGGCT TCAGTTTATT GCTGTATACT CTGTGCTTCT GGCTCTCATG GAAAAGACAG ACATTGGGGT TCTTATAATC TCTCCCTCTC CCCTCCCCAC ACTCTATCCC CAAAGGAGGC ACCACTTCTG CAGGTAAATG TTATCTTCAA AGCGCTCACA TCGCAACCTT TGCCCACACC ATCTCATTAA AGGAATTGGC AGTGACTTTA AGGTGAAAGA ACTCGGTGGC TACGTGTTAT ATAAATTTGC ATCTGGGTCT CAGAGCTGGA AGGAAGGCAC TCCCATACAT GCAGTCTGTA CATGCAGTCG GATGATGGAC CAACAACACA TTGTGATTTA TGCCCCTGCT GGTGAGCCCA GGAATCCCTG TAGCACTCTC TCTCAGCTCT AGGGCCCTGC TTGTGTATGG AAAACGCTTA GTGTTTTATA GGTATTTTGT CAGAATACTT 1 TAAGGAACTT GACCAAAGTT ACAGGGAGGT TAGACAGATT GTCATGGTAT ACTCACCTCT 1 GTCTCTGACC CTCCTAACTG GGACCTCTTT AGTCTCCCTT GAGGCAGGGA GTGCCACATG 1 CATGTGTCCA GGCACATGTC TCCTGGTTTA CCTCCCAACG CACCTCAAGT CCCCAAGGTA 1 GGTAGGCACT TGTATTCTGT AATTCAGAGA GGCAAATCAA ACTGTTACAA TGTTTGCCCA 1 AAGCTCCCCA AGCAAAGTGG CCCTAAGAGT GAGCAAAGAG ACTGCGTGCC TTCACTGCCT 1 GTGTGAATCC CTGCAGATAG TCTCTCATCT TGGTGCCCTT CCCACAGAGG CTGGGGCGGC 1 AGGAGGGAGC CTGGACAGCT CAGACACTGG GTCATTGATG ACTGTTGTGT GGGATACCTG 1 CCGGGGCGCA GGAGTGAGCC ATGCCACCCC AGGAAGTGGT TCAGGGTGAC TCTTCTTGGC 1 ACACCTGGGA GGATGTAGCT GGTGCTGGCA CACCCACCGT CACGAGAGCT TCCTGTCCAA 1 ACCTTCAACA AAGGCGGCTT CTTGAGACAG GCTAGACTGA AGTCACCAGC CTTGGGTGGG 1 GTCCACTATG TAACCTCAGT GCTCAGGAAC CCTTTCCCAT ACTGTCTGGA ACTATACTGT 1 ATGTAGCTGG GTTTCCACGC ATGTGTGCCT GCACCCAGTC CATCTCATCT TCTATCTCCC 1 TCCCCTTTCC CGCTTCCCCC CTCCCCACTC TCCATCTCAT CTTCCATCCC CACCTCTTCT 1 GGTCCCTGCC CTGCTAAACT CAGGGTAGCT GCATTCCGCT GGCCTTCCCC ATGTTCCAGG 1 CTTCAGTCCC TTCTCTGCAC CTGTCCTTTG TGAAGTGACC AGAGGATTTC TGATCCTGTC 1 TCTGTCGCTC TGAAGGGTCA GGAGTTCCTC CTGCCTGGAC AAAGCCATCC TGACGCACAT 1 AAATAAAACA AACATCAAAC TCTATTCAAC CCCCTGGAAC CCGTGTGTGT TACTTACAGG 2 GCAAAAGAAT GGAGCAGGGG ATGGGTTGTG GGGGGGGGGG GTGGCATCTG GGTTGTCTAC 2 AGTTGTGCAT TAAGTTGTAA TTAAGATGTG CATTTCTCCA AATAAGGGAA AATTATTCTG 2 GATTATTTGA GTGAAGCTGA AAGGTGATCA TCTAGA 2

Claims: WHAT IS CLAIMED IS: 1. A method of decreasing the growth of a malignant cell wherein said method comprises exposing said cell to efficacious levels of a compound that induces expression of syndecan in said cell. 2. The method of claim 1 , wherein said induced expression is obtained by affecting the enhancer element of the syndecan gene (SEQ ID No. 3). 3. The method of claim 1 , wherein said induced expression is obtained by affecting the suppressor element of the syndecan gene. 4. The method of claim 1 ,2 or 3, wherein said cell is steroid-responsive. 5. The method of claim 4, wherein said steroid is estrogen or androgen. 6. The method of claim 4, wherein said cell is selected from the group consisting of a malignant breast cell, an endometrium cell and a prostate cell. 7. The method of claim 6, wherein said method further comprises exposing said cell to an anti-steroid agent. 8. The method of claim 7, wherein said agent is toremifene or tamoxifen. 9. The method of claim 1 ,2 or 3, wherein said cell is a mesenchymal cell. 10. The method of claim 6 or claim 9, wherein said cell is a human cell. 11. The method of claim 1 , wherein said cell is exposed to efficacious levels of a composition comprising growth factors of said cell. 12. The method of claim 11 , wherein said growth factors are selected from the group consisting of bFGF; TGF-β; and bFGF and TGF-β. 13. A method for treating a patient in need of treatment to decrease the growth of a malignant tumor in said patient, wherein said method comprises administering, to said patient, efficacious levels of a composition that induces expression of syndecan in the cells of said tumor. 14. A method of claim 13, wherein said composition affects the enhancer element of the syndecan gene. 15. A method of claim 13, wherein said composition affects the suppressor element of the syndecan gene. 16. The method of claim 13, 14 or 15, wherein said tumor is a tumor whose growth is stimulated by steroids. 17. The method of claim 16, wherein said steroid is estrogen or androgen. 18. The method of claim 17, wherein said tumor is selected from a tumor the group consisting of a breast, endometrium, prostate gland or mesenchymal tissue. 19. The method of claim 13, wherein said composition comprises an anti- steroid agent. 20. The method of claim 19, wherein said agent is toremifene or tamoxifen. 21. The method of claim 13, wherein the efficacious agents in the composition comprise growth factors. 22. The method of claim 21 , wherein said growth factor is selected from the group consisting of bFGF; TGF-β; and bFGF and TGF-β. 23. The method of claim 13, wherein said patient is a human. 24. A method of inducing differentiation of a cell to a more differentiated phenotype, wherein said method comprises exposing said cell to efficacious levels of a composition that induces expression of syndecan in said cell. 25. The method of claim 24, wherein the composition affects the enhancer element of the syndecan gene. 26. The method of claim 24, wherein the composition affects the suppressor element of the syndecan gene. 27. The method of claim 24, wherein the growth of said cell is stimulated by steroids. 28. The method of claim 27, wherein said steroid is estrogen or androgen. 29. The method of claim 24, wherein said cell is an epithelium cell. 30. The method of claim 29, wherein said epithelium cell is an epidermal skin cell. 31. The method of claim 24, wherein said cell is a human cell. 32. The method of claim 24, wherein said composition comprises an anti- steroid agent. 33. The method of claim 24, wherein the efficacious agent in said composition is toremifene or tamoxifen. 34. The method of claim 24, wherein the efficacious agent in said composition is a growth factor. 35. The method of claim 34, wherein said growth factor is selected from the group consisting of bFGF; TGF-β; and bFGF and TGF-β. 36. A method of stimulating hair growth in an epidermal cell of skin, wherein said method comprises stimulating expression of syndecan in said cell. 37. The method of claim 36, wherein said stimulated expression is achieved by affecting the enhancer element of the syndecan gene. 38. The method of claim 36, wherein said stimulated expression is achieved by affecting the suppressor element of the syndecan gene. 39. The method of claim 36, wherein said cell is a mesenchymal cell. 40. The method of claim 36, wherein said method comprises administering a composition comprising efficacious levels of a growth factor. 41. The method of claim 40, wherein said growth factor is selected from the group consisting of bFGF; TGF-β; and bFGF and TGF-β.

Inventors: ALANEN-KURKI LEENA MARIA, AUVINEN PETRI OLLI VILJAMI, JAAKKOLA PANU MIKKO, JALKANEN MARKKU TAPANI, LEPPAE SIRPA MARIANNE, MALI MARKKU SAKARI, VIHINEN TAPANI ARTERO, WAERRI ANNI MAIJA, ALANEN-KURKI, LEENA, MARIA, AUVINEN, PETRI, OLLI, VILJAMI, JAAKKOLA, PANU, MIKKO, JALKANEN, MARKKU, TAPANI, LEPPAE, SIRPA, MARIANNE, MALI, MARKKU, SAKARI, VIHINEN, TAPANI, ARTERO, WAERRI, ANNI, MAIJA
Applicants: ALANEN KURKI LEENA MARIA, AUVINEN PETRI OLLI VILJAMI, JAAKKOLA PANU MIKKO, JALKANEN MARKKU TAPANI, LEPPAE SIRPA MARIANNE, MALI MARKKU SAKARI, VIHINEN TAPANI ARTERO, WAERRI ANNI MAIJA, ALANEN-KURKI, LEENA, MARIA, AUVINEN, PETRI, OLLI, VILJAMI, JAAKKOLA, PANU, MIKKO, JALKANEN, MARKKU, TAPANI, LEPPAE, SIRPA, MARIANNE, MALI, MARKKU, SAKARI, VIHINEN, TAPANI, ARTERO, WAERRI, ANNI, MAIJA
Application year: 1993
Main classification: A61K31
IPC codes: A61K31, A61K38, A61K45, A61P35, A61P43, C07K14, C12N5, C12N15, C12R1