bluuebunny/crossref_metadata_2025 huggingface dataset:
An immunohistochemical study of c‐erbb‐2 protein in gastric carcinomas and lymph‐node metastases: Is the c‐erbB‐2 protein really a prognostic indicator?
Authors: Ohguri Takehiko , Sato Yuichi , Koizumi Wasaburo , Saigenji Katsunori , Kameya Toru
Published: February 2007

An immunohistochemical study of the c‐erbB‐2 protein was conducted on formalin‐fixed paraffin‐embedded tissue sections from 136 primary gastric carcinomas and 50 metastatic lymph node tumors obtained at gastrectomy. Expression of the protein was detected in 35 of 136 primary gastric carcinomas (25.7%) and 22 of 50 metastatic lymph nodes (44%). The staining pattern of tumor cells was classified as membranous or cytoplasmic. An immunohistochemical study using serially diluted antibody demonstrated that 82.6% of positive cases in metastatic lymph nodes showed c‐erbB‐2 immunoreactivity stronger than that in the primary tumors. Membranous staining was stronger than cytoplasmic staining. c‐erbB‐2 protein of the cytoplasmic as well as membranous types was confirmed to be a 185‐kDa whole molecule by immunoblotting. Correlation between the expression of c‐erbB‐2 protein and clinical and histological parameters was investigated. No significant correlation between 5‐year survival rate of patients and expression of c‐erbB‐2 protein was found. In the poorly differentiated carcinoma group possessing c‐erbB‐2 protein, overall survival was significantly shorter than in cases without protein expression ( p < 0.01). We conclude that c‐erbB‐2 protein is not a useful prognostic indicator in gastric carcinomas.
Altered glycosylation of β1 integrins associated with reduced adhesiveness to fibronectin and laminin
Authors: Kawano Takehiro , Takasaki Seiichi , Tao Tien‐Wen , Kobata Akira
Published: February 2007

The carbohydrate structures of the β1 integrins obtained from a mouse metastatic melanoma 816 FI and its weakly metastatic wheat‐germ agglutinin‐resistant mutant Wa4‐bI were studied comparatively. The results indicated that the integrins from both cells contain high mannose‐type and bi‐, tri‐ and tetra‐antennary complex‐type sugar chains. No significant difference was found in the outer chain branching between both integrins, but sialylation of the sugar chains of the mutant's integrin was markedly decreased and almost all the outer chain moieties of tri‐ and tetra‐antennary oligosaccharides of the mutant's integrin were fucosylated, resulting in the formation of X‐antigenic determinants, GalβI → 4 (Fucα → 3) GlcNAc. In contrast, the integrin from parental cell contained no X‐antigenic determinant. These structural differences found in the integrin are thought to account for the reduction in the metastatic potential of the mutant which also shows reduced adhesion to fibronectin and laminin as compared with the parental cell.
Biologic properties of a Ch2 domain‐deleted recombinant immunoglobulin
Authors: Slavin‐Chiorini Dale C. , Hand Patricia Horan , Kashmiri S. V. S. , Calvo Benjamin , Zaremba Sam , Schlom Jeffrey
Published: February 2007

Monoclonal antibody (MAb) B72.3 reacts with TAG‐72, a high‐molecular‐weight mucin expressed on several types of human carcinoma, and is currently being used in clinical trials for the diagnosis and therapy of human carcinoma. An expression construct containing cDNA encoding an immunoglobulin (Ig) heavy chain, with the variable region of murine MAb B72.3 and a human Ig constant region with a deletion of the CH2 domain, was generated. Immunoglobulin from the transfectoma with the highest expression of the TAG‐72 immunoreactive antibody was designated MAb chimeric (c) B72.3°CH2. The pharmacokinetics of serum clearance of iodine‐labeled MAbs cB72.3°CH2 and the intact cB72.3 were compared in athymic mice. By 24 hr, less than 1% of the cB72.3°CH2 was left in the plasma, while 36% of the cB72.3 still remained. The T1/2β values of the cB72.3°CH2 and cB72.3 MAbs were 1.7 and 2.4 hr, respectively. The T1/2β values were 7.8 hr for the domain‐deleted cMAb and 48.9 hr for cB72.3. Biodistribution studies in athymic mice bearing LS‐174T xenografts showed a reduction in the percentage of injected dose per gram in tumor with 1311‐cB72.3°CH2; however, the 1311‐cB72.3ΔCH2 both localized to tumors faster and cleared from the blood faster than the 1251‐cB72.3 MAb. Only trace amounts of the 1311‐CB72.3°CH2 were detected in normal tissues, including kidney. The faster clearance rate, more rapid tumor targeting and lack of metabolic uptake in normal tissues demonstrated with the iodine‐labeled CH2 domain‐deleted cMAb may be an advantage for certain clinical protocols.
Interaction of topoisomerase I inhibitors with radiation in cis‐diamminedichloroplatinum(II)‐sensitive and ‐resistantcells In Vitro and in the fsaiic fibrosarcoma In Vivo
Authors: Bosciai Roseanne E. , Korbut Timothy , Holden Sylvia A. , Ara Gulshan , Teicher Beverly A.
Published: February 2007

The cytotoxicity of the topoisomerase‐I inhibitors, camptothecin and topotecan, toward the SCC‐25 human head‐and‐neck squamous‐carcinoma cells and the SCC‐25/CDDP sub‐line made resistant to cis‐diamminedichloroplatinum(II) was assessed alone and in combination with radiation. Topotecan was less cytotoxic than camptothecin in cell culture and the SCC‐25/CDDP cell line was more sensitive to either topoisomerase‐I inhibitor than was the parental SCC‐25 cell line. Both camptothecin and topotecan were effective radiation sensitizers of hypoxic SCC‐25 and SCC‐25/CDDP cells under normal pH or acidic pH conditions. Sensitizer‐enhancement ratios ranged between I.5 and I.6 for hypoxic SCC‐25 cells and between I.3 and I.5 for hypoxic SCC‐25/CDDP cells. When the ability of campothecin or topotecan to sensitize the FSallC fibrosarcoma to single‐dose radiation was assessed using the tumor‐cell‐survival assay, a sensitizer‐enhancement ratio of I.2 was found with each drug. However, using tumor growth delay of the FSallC fibrosarcoma to determine the effect of camptothecin or topotecan to enhance the efficacy of a daily fractionated radiation regimen, topotecan produced a sensitizer‐enhancement ratio of I.4, while that for camptothecin was I.2. These results indicate that topoisomerase‐I inhibitors may retain activity in CDDP‐resistant cells and may be effective adjuncts to radiation therapy.
Macrophage‐induced cytotoxicity and anti‐metastatic activity of a 43‐kDa human urinary protein against the lewis tumor
Authors: Fontan Elisabeth , Saklani Hélène , Fauve Robert M.
Published: February 2007

Resident, inflammatory or bone‐marrow macrophages from C57BI/6 mice incubated in vitro with a pure human urinary protein (HGP.43) decreased the growth rate of Lewis tumor cells (3LL). This inhibition of 3LL growth was the result of a cytotoxic activity of these macrophages which was independent of oxygen metabolites and nitrous oxide. Murine monoclonal antibodies (MAbs) against HGP.43 inhibited macrophage‐mediated cytotoxicity. This cytotoxic activity was not due to the release of cytotoxic factors in the culture supernatant, showing that a contact between macrophages and tumor cells was required to express cytotoxicity. The presence of HGP.43 was absolutely necessary during the incubation of macrophages with target cells. In vivo, in HGP.43‐treated mice, the growth of the primary tumor was not delayed but the size and number of lung metastases were significantly reduced 21 days after tumor inoculation.
Effect of drug‐light interval on photodynamic therapy with meta‐tetrahydroxyphenylchlorin in malignant mesothelioma
Authors: Ris Hans‐Beat , Altermatt Hans Jörg , Nachbur Bernhard , Stewart J. Charles M. , Wang Qiang , Lim Chang Kee , Bonnett Raymond , Althaus Ulrich
Published: February 2007

The influence of the time interval (TI) between drug administration and laser activation on selectivity of meta‐tetrahydroxy‐ phenylchlorin(mTHPC)‐mediated photodynamic therapy (PDT) for tumour tissue was assessed in BALB/c nude mice bearing human malignant mesothelioma xenografts. Following i.p. administration of 0.3 mg/kg mTHPC, a light dose of 10 J/cm2 and 0.1 W/cm2 was delivered at 650 nm on the tumour and an equal‐sized area of the hind leg after 4. I2, 24 and 36 hr and 2,3, 4,5 and 6 days to groups of 6 animals (surface irradiance). Then, 72 hr after light delivery, the depth of necrosis was measured in the tumour and in the skin and underlying muscle of the hind leg. Photosensitized necrosis occurred in normal tissue at TI from 4 hr to 3 days and in the tumour at TI from I2 hr to 4 days. The therapeutic ratio of mTHPC‐PDT varied significantly with the time interval between drug administration and laser activation and was greatest at an interval of 3 days. mTHPC concentration was measured in 3 control unirradiated animals at all time points in normal tissues and in tumour tissue, and found to be the same in both tissues. Thus the tissue concentration of mTHPC was of limited use as regards the prediction of photosen‐sitizing effects in the tumour model.
Epstein‐barr virus (EBV) nuclear‐antigen‐2‐induced up‐regulation of cd21 and cd23 molecules is dependent on a permissive cellular context
Authors: Cordier‐Bussat Martin , Billaud Marc , Calender Alain , Lenoir Gilbert M.
Published: February 2007

The Epltein‐Barr virus (EBV) induces unlimited growth of B lymphocytes in vitro, a phenomenon known as immortalization. The elucidation of the mechanisms by which EBV de‐regulates 8‐cell protiiferation in vitro will permit an undemtanding of how the vim contributes in vivo to the genesis of Burkitt's lym‐ phoma (BL) and of lymphopmliierations in immunosuppressed patients. At present, no singie EBV immortalizing gene has been identified, and the hypothesis has been made that many viral genes cooperate in establishing an autocrine loop of secretion leading to irnmortdization. Constitutive expression of B‐cell surface molecules such as CD2l and CD23, specifically impli‐ cated in the control of B‐cell proliferation, is indeed induced at the surface of immortalized B lymphocytes. The expression of the v i d nuclear antigen 2 (EBNA2) has been shown to be in part responsible for CD2l and CD23 upregulation, and EBNA2 is suspected to be a transactivahr of cellular genes, although this point remains to be demonstrated. The role of EBNA2 gene, independently of other vim1 genes, has beeriinvestigated by transfeetion into B‐lymphoma lines, but conflicting results have been reported. To further investigate its role in the regulation ofCD2l and CD23 molecules, we have compared the effects of EBNA2 expression in 2 sets of B‐lymphoma lines infected with P3HRI EBV strain, and/or tmnsfected with EBNA2 gene. We report here that: (i) EBNAZ expression is not a sufficient conditiw to induce CD21 and CD23 upregulation, EBNA2′s effects are highly dependent on the cellular context, and momover can be modffied by infection with P3HRl virus; (ii) EBNA2 inducer acthation of CD23 expression in a very pvticutar way, namely, an increased quantity of CD23 steady‐ state RNA coding for the form A of the protein, which is not detectable at the cell surface but directly secreted. ® 1993 Wiley‐Liss, Inc.
Identification of T‐ and B‐cell epitopes associated with a restricted component of the epstein‐barr virus‐induced early antigen complex
Authors: Pothen Susan , Cao Tin , Smith Richard , Levine Paul H. , Levine Alexandra , Pearson Gary R.
Published: February 2007

Experiments were designed in an attempt to identify T‐and B‐cell epitopes expressed on the 17‐kDa early‐antigen‐restricted (EA‐R) polypeptide of the EBV‐induced early antigen complex. Using Berzofsky's algorithm, 3 hypothetical T‐cell epitopes on p 17 were synthesized and employed in EBV‐specific lymphoproliferative assays. Lymphocytes from all EBV‐infected donors responded against one of these epitopes (p 17.1) irrespective of their serological status relative to antibodies to EA‐R. Both CD4+ and CD8+ T‐cell subpopulations from seropositive donors proliferated in the presence of p17.1 in short‐term cultures. These experiments therefore identified one T‐cell epitope on the 17‐kDa polypeptide. In contrast, sera from anti‐EA antibody‐positive individuals reacted with all 3 synthetic peptides to varying degrees, with p 17.1 being the most frequently reactive epitope. When the sera were grouped according to diagnosis, it was noted that 82% of the sera from patients with aggressive lymphomas, whether Africans with Burkitt's lymphoma or North Americans with intermediate‐grade large‐cell or high‐grade B‐cell lymphoma, contained antibody reactive with p17.1, while 64% were reactive with p 17.2 and 29% with p 17.3. In contrast, high anti‐EA antibody‐positive sera from nasopharyngeal carcinoma patients were relatively less reactive with these synthetic peptides (23% positive with p17.1; 19% with p17.2; and 13% with p17.3). These results therefore identified 3 8‐cell EA‐R epitopes which might be potentially useful for clinical or epidemiological studies of EBV‐associated lymphoproliferative diseases.
The value of the 1981 who histological classification in inter‐observer reproducibility and changing pattern of lung cancer
Authors: Campobasso Onofrio , Andrion Alberto , Ribotta Marisa , Ronco Guglielmo
Published: February 2007

A series of 722 lung carcinomas, surgically resected and typed some years ago according to the 1967 WHO classification, was independently reviewed by 2 observers in order to test the reproducibility of histopathological typing when using the criteria of the 1981 WHO classification. Typing was fully agreed upon in 87% of cases. Agreement was very high for squamous‐cell, small‐cell and adeno‐carcinomas (K = 0.87,0.89 and 0.85, respectively) while adenosquamous (K = 0.56) and large‐cell (K = 0.71) carcinomas were more controversial categories. A consensus diagnosis was formulated for lesions with discrepant diagnoses. When comparing the final typing to the previous typing based on the 1967 WHO classification, squamous‐and large‐cell carcinomas were reduced respectively by 22% and 33% and adenocarcinomas increased by 94% of the original number. The 1981 “adenosquamous carcinoma” category included 2.8% of the tumours, while the corresponding 1967 category “combined epidermoid and adenocarcinoma” was empty. These changes must be taken into account when considering epidemiological studies, especially those aiming at evaluation of the secular trends of lung cancer by cell type.
Site distribution of cutaneous melanoma in queensland
Authors: Green Adèle , Maclennan Robert , And Phillipa Youl , Martin Nicholas
Published: February 2007

The age‐adjusted incidence rates of cutaneous melanoma in Queensland in 1987 have been analysed for 16 anatomic sites, taking into account their surface areas. In men, the incidence of invasive melanoma on the ears, a chronically sun‐exposed site, was extraordinarily high with annual rates of over 200 per 105 units of surface area in the Queensland population. Next highest rates of over 100 melanomas per 105 units were found on the face, neck, shoulders and back in men and the face and shoulders in women. Comparison with site‐specific incidence rates in the same population 7½ years previously showed that incidence of invasive disease had significantly increased for all these sites, though the largest relative increase in this period occurred on the forearm in both men and women. Melanoma was very rare on the buttocks of both sexes and on the scalp in women, sites which receive the least sun exposure. These findings are consistent with the theory that excessive total sun exposure plays a major role in the aetiology of cutaneous melanoma.
Sequence variation in the E7 gene of human papillomavirus type 18 in tumor and non‐tumor patients and antibody response to a conserved seroreactive epitope
Authors: Ter Meulen J. , Schweigler A. C. , Eberhardt H. C. , Luande J. , Mgaya H. N. , Müller M. , Bleul C. , Ulken V. , Ikenberg H. , Pawlita M. , Gissmann L.
Published: February 2007

We have determined nucleotide sequences of the E7 open reading frame (ORF) of human papillomavirus type 18 (HPV‐18) isolates obtained from 18 cervical carcinomas from Tanzanian and German patients and 8 cervical scrapings from Tanzanian non‐tumor patients. The HPV‐18 prototype sequence was detected in only 3 out of 26 isolates. Silent mutations were found at nt positions 640 and 751, whereas the mutations observed at nt positions 770, 806, 864 and 865 all change the respectively encoded amino acid. The HPV‐18 isolates of 3 German carcinomas showed the same mutations (at position 751) as those of 2 established cervical carcinoma cell lines (HeLa and C4‐1), whereas different mutations were found in 16/23 African isolates (at positions 640 and 864), to which the isolate of cell line SW756 was similar (changes at positions 640 and 865). Seven out of 15 HPV‐18‐positive Tanzanian tumor patients (46.7%) reacted in a peptide ELISA against a recently described seroreactive epitope of the HPV‐18 E7 ORF (nt positions 704‐769). Mutational changes of the E7 ORF were excluded as a possible explanation for the lack of antibody response, because there was no correlation with the serological results. The seroreactive region appears to be well conserved despite geographically varying mutations within the E7 ORF of HPV‐18.
Myoepithelial and basement membrane antigens in benign and malignant human breast tumors
Authors: Guelstein V. I. , Tchypysheva T. A. , Ermilova V. D. , Ljubimov A. V.
Published: February 2007

Serial cryostat sections of 160 human breast lesions and of 9 lymph‐node metastases were studied by indirect immunofluores‐cence. We used monoclonal antibodies (MAbs) to lining‐epithelium‐specific keratin 8 and to myoepithelium‐specific keratin 17 in combination with polyclonal and monoclonal antibodies to major basement membrane components, laminin, collagen type IV, entactinhidogen, and large heparan sulfate proteoglycan (perlecan) core protein. Continuous basement membranes adjacent to a basal layer of keratin‐17‐positive myoepithelial cells were typical for normal, benign and in situ carcinomatous structures. In invasive and metastatic structures, always formed by keratin‐8‐positive tumor cells, basement membranes were found only rarely and with conspicuous fragmentations. This lack of basement membranes correlated with loss of myoepithelium identified by staining for keratin 17. In comedo structures of invasive ductal carcinomas and in papillary carcinomas, fibrovascular complexes with numerous blood vessels and deposition of basement membrane material were often seen in the stroma. lmmunomorphological analysis of 41 cases of doubtful diagnosis at intra‐operative biopsy was also performed. A combination of MAbs to keratins 8 and 17, and to basement membrane components, made it possible to distinguish between morphologically similar benign and malig‐nant proliferations and to detect single‐cell invasion of the stroma. This combination of antibodies may be recommended as an auxiliary immunomorphological tool for differential diagno‐sis of intra‐operative breast biopsies in dubious cases.
Expression of functionally intact pdgf‐α receptors in highly metastatic 3ll lewis lung carcinoma cells
Authors: Fitzer‐Attas Cheryl , Feldman Michael , Eisenbach Lea
Published: February 2007

The ability of disseminating tumor cells to grow in a target organ is the final limiting step of the metastatic cascade. The growth of a highly lung‐metastatic clone, D122, of the murine 3LL Lewis lung carcinoma was induced in vitro with lungconditioned media (CM) to a greater extent than that of a weakly metastatic clone, A9. With the use of platelet‐derived growth factor (PDGF)‐α‐receptor‐specific antibodies, the possible paracrine mode of metastatic cell growth was further suggested by demonstrating the presence of these receptors on highly metastatic cells only. Receptors for PDGF have been found almost exclusively on cells of mesenchymal and glial origin. Therefore, the functionality of this receptor in the D122 epithelial cell line was verified by immune complex kinase and ligand‐stimulation assays. Moreover, PDGF was shown to specifically induce in vitro growth of D122 highly metastatic cells only and to be abundantly expressed in lung CM but not in kidney or liver CM. Thus, the interaction of PDGF‐like factors in the lung with PDGF receptors on the metastatic tumor cell may be important in the development of metastatic lesions in the target organ.
Hypersensitivity of human testis‐tumour cell lines to chemotherapeutic drugs
Authors: Masters John R. W. , Osborne Edward J. , Claire Walker M. , Parris Christopher N.
Published: February 2007

Metastatic testis tumours, in contrast to most other types of cancer, can be cured by drugs. To investigate which classes of chemotherapeutic drug are differentially toxic to testis‐tumour cells, we compared the in vitro dose‐response curves of 5 human testis and 5 bladder‐cancer cell lines to 12 compounds. The testis cells were hypersensitive to drugs that interact directly with DNA (m‐amsa, bleomycin, cisplatin, doxorubicin, methylni‐ trosourea, mitozolomide, etoposide, mitomycin‐C), but little or no difference between the 2 cell types was seen following exposure to drugs whose mechanisms of action do not involve direct interaction with DNA (methotrexate, 5‐fluorouracil, colchicine, vinblastine). We conclude that testis tumour cells are either less tolerant of, or have a reduced capacity to repair, DNA damage.
International variations in the incidence of childhood bone tumours
Authors: Parkin D. M. , Stiller C. A. , Nectoux J.
Published: February 2007

Bone cancers comprise about 5% of childhood neoplasms. Osteosarcoma, the most common sub‐type, shows a somewhat irregular geographic pattern of incidence, with low rates in some Asian (Indian, Japanese, Chinese) and Latin American populations. Incidence is similar in the sexes and rises steeply with age, accompanied by an increasing proportion of tumours localized in the long bones of the legs. Rates in the USA are higher in blacks than in whites, as a result of a higher incidence at ages 10 to 14 and of tumours of the leg bones. The descriptive epidemiology is consistent with early observations linking risk to the amount of bone growth. Ewing's sarcoma is rare in black populations (USA and Africa) and in eastern Asia. Compared with osteosarcoma, a lower percentage of tumours is localized to the long bones, and incidence rises less steeply with age and is accompanied by an increasing proportion of pelvic tumours. Chondrosarcoma is a rare cancer in children (less than 5% of bone cancers), with an age distribution similar to that of osteosarcoma and a sub‐site distribution resembling that of Ewing's sarcoma. Little is known of the aetiology of these tumours; there is clearly a strong genetic predisposition in Ewing's sarcoma but, although the proportion of osteosarcorma cases of genetic origin seems to be small, environmental determinants so far suspected can account for only a small fraction of the total cases.
Analysis of the relationship between stage of differentiation and nk/lak susceptibility of colon carcinoma cells
Authors: Blottière H. M. , Zennadi R. , Grégoire M. , Aillet G. , Denis M. G. , Meflah K. , Le Pendu J.
Published: February 2007

NK and LAK cells which are able to lyse tumor target cells in an MHC‐unrestricted manner are not equally effective against targets of the same nature. In the case of colorectal tumors, some cells are highly sensitive, whereas others are resistant to NK and can even be quite resistant to LAK‐mediated lysis. In the present paper, we tried to correlate the stage of differentiation of 17 human colorectal tumor cell lines with their NK‐ or LAK‐cell susceptibility. It was observed that NK cells killed colorectal target cells independently from their stage of differentiation defined according to histopathological criteria from xenografting in nude mice. NK susceptibility was not correlated either with in vitro‐defined criteria of differentiation, such as cell polarity and morphology, brush‐border enzyme expression and CEA production. A LAK‐resistant HT‐29 sub‐line (HT‐29 LAK) was selected which could not be distinguished from HT‐29 in terms of features of differentiation. It was further observed that HT‐29 Glc−/+ cell line, a highly differentiated enterocytic‐like variant of HT‐29, obtained after glucose starvation, was killed by LAK cells as efficiently as the moderately differentiated parental HT‐29, and that Caco‐2 cells, which differentiate spontaneously after confluence in standard culture conditions, were equally sensitive to NK‐mediated lysis whatever their stage of differentiation. In contrast, HT29 MTX10−5, a highly differentiated mucus‐secreting variant of HT29 obtained by methotrexate selection, was much more resistant to LAK cells than parental HT29 cells.
Monocyte‐conditioned media possess a novel factor which increases motility of cancer cells
Authors: Jiang W. G. , Puntis M. C. A. , Hallett M. B.
Published: February 2007

A number of cytokines have been reported to increase the cell motility (scatter) of cohesive cell colonies. We report here a novel scattering factor produced by human monocytes. Media from both stimulated and non‐stimulated human monocytes or the monocytic cell line, U937, increased the cell motility of 2 human colon‐cancer cell lines, HT115 and HT29, but not the canine epithelial cell line MDCK. Motility was assayed by cell dissociation from carrier beads and colony scattering. HT115 cells were strongly scattered by the conditioned media but not by interleukins IL‐1,2,4,6,8,10, TNFα, TGFβ, EGF, GM‐CSF, IGF‐1, PDGF, interferon‐γ. This factor is distinct from hepatocyte growth factor/scatter factor (HGF/SF), since the activity was not blocked by anti‐HGF/SF antibody. The activity was reduced by treatment with acid, heat, trypsin and dithiothreitol; this, together with gel filtration, suggests that the factor is a protein (MW 40 to 60 kDa). This new factor, which is secreted from monocytes and the monocytic cell line, U937, has the ability to increase the motility of cancer cells and may be important in controlling the behaviour of tumours in vivo.
Expression and distribution of peripherin protein in human neuroblastoma cell lines
Authors: Pedersen Ward A. , Becker Laurence E. , Yeger Herman
Published: February 2007

A series of human neuroblastoma (NB) cell lines was analyzed for expression of peripherin, a class‐III intermediate filament protein expressed at high levels in ganglia of the peripheral nervous system. By Western blotting, peripherin protein was detected in all human NB cell lines examined. The highest level of peripherin was found in the NUB‐7 cell line, previously characterized as homogeneously neuroblastic. By immunofluorescence labeling, peripherin was shown to be organized in a perinuclear filamentous pattern and, exemplified by IMR32 cells, was also shown to be localized to spontaneously formed neurites. Peripherin was expressed in neuroblastic but not substrate‐adherent cells, and was found at low levels in I‐type cells. There was a pronounced redistribution of peripherin to neurites formed in response to dibutyryl cyclic adenosine monophosphate (dbcAMP) and all‐trans‐retinoic acid (RA). In NUB‐7 cells, which do not extend neurites in response to nerve growth factor, there was no change in the level of peripherin protein following treatment with this agent. Both dbcAMP and RA induced a redistribution of peripherin to neurite extensions, but only treatment with RA increased the level of the protein as demonstrated with NUB‐6A4 and NUB‐6C4 subclones. Peripherin was also variably expressed in peripheral neuroepithelioma (NE) cell lines tested, but was organized into a more basket‐like filamentous pattern in these cells. The heterogeneous expression and distribution of peripherin in NB and NE cell lines indicate that this protein is associated with maturation of the neuronal phenotype and hence serves as a differentiation marker for tumors derived from the neural crest.
Anti‐metastatic vaccination of tumor‐bearing mice with il‐2‐gene‐inserted tumor cells
Authors: Porgador Angel , Gansbacher Bernd , Bannerji Rajat , Tzehoval Esther , Gilboa Eli , Feldman Michael , Eisenbach Lea
Published: February 2007

IL‐2 gene was introduced through retroviral vectors into the highly malignant and poorly immunogenic 3LL‐D122 clone. Both high and low D122‐11‐2 secretors showed elimination of tumorigenicity in syngeneic immune‐competent mice; however, in nude mice only the high IL‐2 secretor showed reduced tumorigenicity as compared with parental D 122 cells. Also, following intravenous inoculation, only the high IL‐2 secretor showed reduced generation of metastases, whereas the low IL‐2 secretors were as highly metastatic as parental cells. These results seem to indicate that low levels of IL‐2 secreted by tumor cells are sufficient to activate T cells, while higher levels are needed in order to activate non‐T‐cell effectors. D122‐IL‐2 secretors induced high levels of anti‐tumor CTL, while their sensitivity to the lytic activity of these CTL was similar to the sensitivity of D122 cells, thus indicating that the elevation of immunogenicity of IL‐2 secretors was essentially due to the secreted IL‐2. In accordance with CTL induction, pre‐immunization with IL‐2 secretors protected mice against subsequent challenge of parental cells. Moreover, immunization in an “immunotherapy protocol” i.e., vaccination of mice carrying an established tumor of parental D122 cells with inactivated D122‐IL‐2 infectants, almost completely eliminated the generation of lung metastases by D122 cells, thus providing a rationale for the use of IL‐2 gene transferred tumor cells as a modality for treatment of metastasis.
Cellular and In Vivo characterization of the MCR rat mammary tumor model
Authors: Gutzwiller Alois , Regenass Urs , Matter Alex , Lydon Nicholas B.
Published: February 2007

The slowly growing, transplantable MCR‐83 rat mammary tumor is estrogen‐dependent and non‐metastasizing. A rapidly growing, estrogen‐independent, metastasizing subline (MCR‐86) was subsequently isolated in vivo. We have established and characterized cell lines from both MCR rat mammary tumors. MCR cell lines and tumors were studied in vivo and in vitro. Analysis of DNA from tumors and cell lines showed that mutations had not occurred in codons 12, 13 and 61 of the Ha‐ras and Ki‐ras genes. Additionally, dominant transforming activity could not be detected by DNA transfection using NIH 3T3 focus‐forming assay. No gene amplification was detected for either the EGF‐receptor or c‐erbB‐2 genes. Differences in the tyrosine phosphorylation patterns were found between the 2 MCR cell lines. Addition of serum to starved cells resulted in the tyrosine phosphorylation of a 120‐kDa protein, which was elevated in the MCR‐86. The lack of ras activation in the MCR tumors differentiates this model from the widely studied, chemically induced rodent mammary tumors. In addition, the differences in the cellular phosphotyrosine patterns between MCR‐83 and MCR‐86 suggests the occurrence of alterations in signalling pathways that involve tyrosine protein kinases.
Co‐amplification of c‐myc/pvt‐1 in immortalized mouse b‐lymphocytic cell lines results in a novel pvt‐1/aj‐l transcript
Authors: Huppi Konrad , Siwarski David , Shaughnessy John D. , Frederic Mushinski J.
Published: February 2007

In a series of mouse pvt‐I cDNA clones prepared from an immortalized B‐cell line that contains an amplified c‐myc/pvt‐I region, we have identified a unique cDNA, AJ‐I, which contains 57 bp of pvt‐I sequence (pvt‐la) spliced to a novel sequence of 2.1 kb. We report here that this 3′ segment (termed AJ‐IX) maps more than 60 kb telomeric to pvt‐la and is encoded by novel locus (AJ‐I) on mouse chromosome 15. The AJ‐IX probe detects a transcript that is expressed only in normal mouse brain and testis. Several mast‐cell tumors, Ly‐I+ B‐lymphocytic cell lines and a neuroblastoma also display abundant levels of AJ‐IX‐specific mRNA. However, splicing of pvt‐la to AJ‐IX found exclusively in Ly‐I+ B‐cell lines that contain amplified c‐myc/pvt‐I. We conclude that some feature in the generation of this amplicon facilitates the synthesis of a pvt‐I/AJ‐IX chimeric mRNA that may play a role in immortalization of these Ly‐I+ B‐cell lines.
Persistence of plasmin‐mediated pro‐urokinase activation on the surface of human monocytoid leukemia cells In Vitro
Authors: Tapiovaara Hannele , Stephens Ross W. , Vaheri Antti
Published: February 2007

Human leukemia cell lines, unlike those from adherent tumors, have been shown to continuously activate the prourokinase (pro‐u‐PA) they produce. In the present study we found that, in normal cell‐culture conditions in 10% FCS the plasminogen activation cascade works continuously on monocytoid leukemia cells, which expressed plasmin activity and active u‐PA on their cell surface. This plasmin catalyzed the conversion of the produced pro‐u‐PA to active 2‐chain urokinase (tcu‐PA), and was derived from bovine serum plasminogen the activity of cell‐bound tcu‐PA. Plasmin generation was abolished and pro‐u‐PA accumulated in cell cultures that were grown for several days, either in the presence of serum thoroughly depleted of plasminogen, or in the presence of I mM tranexamic acid. Plasmin generated on the cell surface was found to be present in 2 enzymatically active fragments, of 85,000 and Mr 50,000, which were slowly released into the growth medium. These fragments could activate pro‐u‐PA serum‐free growth medium. Most of the bound plasmin could be washed off cells with 10 mM tranexamic acid, but complete removal of plasmin from the cell surface required washing of the cells with acid‐glycine pH 3.0.
Action of a cd24‐specific deglycosylated ricin‐a‐chain immunotoxin in conventional and novel models of small‐cell‐lung‐cancer xenograft
Authors: Zangemeister‐Wittke Uwe , Lehmann Hans‐Peter , Waibel Robert , Wawrzynczak Edward J. , Stahel Rolf A.
Published: February 2007

The therapeutic efficacy of an immunotoxin, SWA II‐SPDB‐dg.ricin A chain, recognizing the leukocyte‐differentiation antigen CD24, was evaluated against SCLC cell lines in tissue culture and in 2 nude‐mouse models. The first model used conventional S.C. solid‐tumor xenografts. The second used small tumor‐cell deposits established in S.C. implanted sponge matrices and allowed us to directly estimate the killing efficiency of the immunotoxin under experimentally defined conditions in vivo. It also mimics the clinical setting of disseminated tumor cells which form the basis of residual disease in SCLC. The cytotoxic potency of SWAII‐SPDB‐dg.ricin A chain was demonstrated in tissue culture by the inhibition of 3H‐leucine incorporation and by the selective elimination of CD24‐positive tumor cells in clonogenic assays. In nude mice, SWAII‐SPDB‐dg.ricin A chain was cleared from the blood circulation with biphasic kinetics: an initial (α phase of 1 hr and a second β phase of 20.5 hr. Following i.v. injection of a dose equivalent to 30% of the LD50, the immunotoxin delayed the growth of SW2 solid‐tumor xenografts by 16 days. The therapeutic efficacy of SWAII‐SPDB‐ dg.ricin A chain was further demonstrated by the selective elimination of clonogenic SW2 cells from small tumor‐cell deposits established in sponge matrices. Regrowth of the solid tumors after the initial response and the clonogenic activity in the sponge‐derived cell population were mediated by CD24‐positive cells, excluding the selection of CD24‐negative mutants during immunotoxin therapy.
Accumulated allelic losses in the development of invasive urothelial cancer
Authors: Habuchi Tomonori , Ogawa Osamu , Kakehi Yoshiyuki , Ogura Keiji , Koshiba Masahiro , Hamazaki Shuji , Takahashi Rei , Sugiyama Taketoshi , Yoshida Osamu
Published: February 2007

To investigate the roles of allelic loss in the development of urothelial cancer, loss of heterozygosity was examined on 7 chromosomal arms in 49 cases of urothelial cancer of various grades and stages. Loss of heterozygosity was found on alleles in order of frequency as follows: 9q (21/38, 55%), 11p (20/44, 45%), 17p (18/42,43%), 13q (10/39,26%), 3p (8/41, 20%), 10q 2/29, 7%) and 1p (1/36, 3%). lnvasive (high‐grade or ≥pT2) tumors showed the loss of 17p (13/16,81%) and the loss of 13q (7/16, 44%) with significantly higher frequencies than non‐invasive (grade 1‐2 ≤pTI ) tumors. Although the loss of 3p and the loss of 11p were also more frequently associated with the invasive phenotypes, the loss of 11p was detected in a considerable number (9 of 26,35%) of non‐invasive tumors. Our results indicate that the loss of 11p might generally occur at an earlier stage before the loss of 3p, 13q or 17p in tumor progression. Since no correlation was found between the loss of 9q and the tumor grade or stage, this genetic alteration appears to be unrelated to invasiveness, and could be one of the initial events in tumorigenesis. Although accumulated allelic losses of 3p, 11p, 13q and 17p are considered to be involved in the development of the invasive type of urothelial cancers, these multiple genetic alterations may have already occurred in some pathologically non‐invasive urothelial cancers. Furthermore, there appears to be some variation in the pattern of cumulative allelic loss.
Mother‐to‐child transmission of human T‐lymphotropic virus type I (HTLV‐I): An extended follow‐up study on children between 18 and 22‐24 years old in Okinawa, Japan
Authors: Ueda Kohji , Kusuhara Koichi , Tokugawa Ken , Miyazaki Chiaki , Okada Kenji , Maeda Yoshiaki , Shiraki Hiroshi , Fukada Kenji
Published: February 2007

The significant difference observed between the seroprevalence of HTLV‐I in adults and in children is as yet unexplained. To evaluate a hypothetical explanation of the existence of seroconversion cases of „seronegative carriers”︁ for this phenomenon, 21 of 55 children who had been born to seropositive mothers and who remained seronegative until the age of 18 years were further followed up at the ages of 22 and/or 24 years. None of the 21 seronegative children born to seropositive mothers seroconverted, either at 22 years or at 24 years. In addition, the polymerase‐chain‐reaction (PCR) technique could not prove the existence of the HTLV‐I provirus genome in peripheral mononuclear cells (PBMC) of 10 of these children. Our results fail to prove the possibility of viral latency of HTLV‐I in mother‐to‐child transmission. Therefore, the hypothetical seroconversion of „seronegative carriers”︁ after adulthood can not be an explanation.
Down‐regulation of integrated Epstein‐Barr virus nuclear antigen 1 and 2 genes in a burkitt lymphoma cell line after lymphoblastoid cells somatic cell fusion with autologous EBV‐immortalized
Authors: Wolf Jurgen , Pawlita Michael , Klevenz Britta , Frech Barbara , Freese Ulrich‐Karl , Müller‐Lantzsch Niko , Diehl Volker , Hausen Harald Zur
Published: February 2007

Epstein Barr virus (EBV) latent gene expression was analyzed in somatic cell hybrids between an EBV‐positive Burkitt lymphoma (BL) cell line (BL 60) and an autologous EBV‐immortalized lymphoblastoid cell line (LCL, IARC 277). The EBV genomes carried by the parental cell lines differ in sequence and in their physical state. The BL 60 EBV genome is integrated into the host cell genome whereas the LCL IARC 277 carries exclusively episomal EBV molecules. The hybrid cells contain both EBV genomes and display the differentiation phenotype of the parental LCL with regard to growth characteristics and cell‐surface antigen expression in vitro and in vivo. While the EBNA 1 and EBNA 2 gene expression of the LCL‐derived EBV is maintained in these hybrid cells, the BL‐60‐derived EBNA 1 and EBNA 2 genes are transcriptionally down‐regulated. Mapping of the genomic region surrounding the latent Cp promoter of the BL‐60‐derived EBV revealed an extensive deletion upstream of the Cp promoter including the enhancer element in the ori P region, the origin of latent viral replication (ori P), the coding sequences for the EBV latent membrane protein (LMP) and the EBV terminal protein (TP), and suggested that one viral‐cellular junction sequence is located near the Cp promoter. Integration of EBV into the host cell genome together with the extensive deletion might be causally related with the altered latent gene expression pattern after introduction of a lympho‐blastoid host‐cell background by somatic cell fusion. Down‐regulation of the BL‐60‐derived EBNA genes could be due to loss of regulatory sequences in the BL‐derived EBV necessary for EBNA 1 and EBNA 2 transcription in the lymphoblastoid hybrid cells, but not in the parental BL cells.
Molecular and cellular heterogeneity of wilms' tumor
Authors: Velasco Susana , D'Amico Domenico , Schneider Nancy R. , Timmons Charles , Chappell Elizabeth , Lee Daniel , Nisen Perry D.
Published: February 2007

We developed a Wilms' tumor‐cell culture system to investigate the molecular basis of nephrogenesis and oncogenesis. Several distinct fractions of cells were isolated and characterized from the same tumor specimen. The cells exhibited striking differences in morphology, immunocytochemical staining profiles and cytogenetics. One fraction contained cells with features of epithelium; other cell fractions resembled partially differentiated mesenchyme (blastema or stroma). While the Wilms' tumor‐suppressor gene WTI was not altered, loss of heterozygosity (LOH) and an insertion in intron I of the p53 tumor‐suppressor gene occurred in the tumor and the cultured cell types. LOH for RB was detected only in the cultured cells. These findings are consistent with a model of tumor initiation in a pluripotent cell that is able to undergo subsequent differentiation along multiple different lines and which mimics normal nephrogenesis.
Production of interleukin‐1β and interleukin‐6 in hepatoblastoma
Authors: Schweinitz Dietrich Von , Hadam Martin R. , Welte Karl , Mildenberger Hermann , Pietsch Torsten
Published: February 2007

Thrombocytosis and fever are frequent symptoms in children with hepatoblastoma. Interleukin‐6 (IL‐6) has been shown to mediate thrombocytosis and an acute‐phase reaction including fever. We therefore investigated samples from I4 untreated patients with hepatoblastoma for this cytokine and in addition for interleukin‐I α (IL‐Iα), interleukin‐Iβ (IL‐Iβ) and tumor necrosis factor‐α (TNF‐α), all of which are known to induce IL‐6 production. High serum levels of IL‐6 were only found in 3/I4 patients; the other cytokines were not detectable. In contrast, I2/I4 tumors produced substantial amounts of IL‐6 in primary cell culture, while IL‐Iβ was found in 3/I4 supernatants; IL‐Iα and TNF‐α were always negative. Immunoenzymatic staining of fresh tumors revealed that IL‐6 is not produced by the tumor cells, but rather by surrounding fibroblasts and endothelial cells. In tumor cells only IL‐Iβ, but neither IL‐Iα, TNF‐α nor IL‐6, could be detected. In co‐culture experiments with fibroblasts and endothelial cells, addition of hepatoblastoma cells enhanced IL‐6 production. Including an IL‐I receptor antagonist abolished this effect incompletely. Our results suggest that tumor cells in hepatoblastoma induce IL‐6 production in surrounding fibroblasts and endothelial cells by virtue of their endogenous secretion of IL‐Iβ and supposedly some other, as yet unidentified, mediator.
Cholecystectomy and colorectal cancer: Evidence from a cohort study on diet and cancer
Authors: Goldbohm R. Alexandra , van den Brandt Piet A. , van't Veer Pieter , Dorant Elisabeth , Sturmans Ferd , Hermus Rudolph J. J.
Published: February 2007

The association between cholecystectomy and subsequent risk for colorectal carcinoma was investigated in a prospective cohort study on diet and cancer (n = 120,852), which is being conducted in the Netherlands from 1986 onwards among 120,852 men and women, aged 55 to 69. After a follow‐up period of 3.3 years, 478 incident cases of colorectal cancer (258 men and 220 women) were identified in the cohort, 64 of whom reported at baseline to have undergone previous gall‐bladder surgery (21 men and 43 women). After adjustment for age and familial history of large‐bowel cancer, the relative rate (RR) for colorectal cancer in subjects who had undergone cholecystectomy compared with those who had not was 1.81 in men (p=0.02) and 1.47 in women (p=0.05). Additional adjustment for parity, Quetelet index, alcohol intake and other dietary variables resulted in a RR of 1.78 in men and 1.51 in women. In women, the highest RR was detected in the right colon (RR = 1.89), whereas in men, no site within the large bowel accounted specifically for the increased relative rate. In both men and women, the rate appeared to increase from approximately 6 years after cholecystectomy onward. According to the TNM stage of the disease, cholecystectomized patients were not detected at an earlier stage than the other patients. It is concluded that in this study the positive association between colorectal cancer and cholecystectomy cannot be explained by detection bias or ascertainment bias and is not confounded by risk factors for gallstone disease or dietary factors.
Orthodox jews have a lower incidence of malignant melanoma. A note on the potentially protective role of traditional clothing
Authors: Vardi Gil , Modan Baruch , Golan Rachel , Novikov Ilya , Shafir Raphael
Published: February 2007

The incidence of malignant melanoma (MM) in orthodox and non‐orthodox Jews in Israel during the years 1970‐1982 was compared through an assessment of incidence rates in the orthodox and non‐orthodox neighborhoods in the city of Jerusalem and by a comparison of the rates in 2 cities on the outskirts of Tel Aviv, one of which has a strongly orthodox population. Within the city of Jerusalem the incidence of MM for the European‐American born in the orthodox neighborhoods was significantly lower than in the rest of the city in males, and of borderline significance in females. A similar pattern was seen upon comparison of the orthodox city of Bnei Brak to its neighbor city, Givatayim. The findings are consistent with the role of sun exposure in the etiology of malignant melanoma.
Relation between autoradiographically measured blood flow and ATP concentrations obtained from imaging bioluminescence in tumors following hyperthermia
Authors: Dellian Marc , Walenta Stefan , Kuhnle Gerhard E. H. , Gamarra Fernando , Mueller‐Klieser Wolfgang , Goetz Alwin E.
Published: February 2007

The effects of moderate local hyperthermia (43.3°C/30 min) on regional blood flow and regional ATP distribution in the amelanotic hamster melanoma A‐Mel‐3 were investigated by high‐resolution techniques. Blood flow and ATP concentrations were measured simultaneously in treated and untreated tumors and in adjacent tissues by means of (14C)‐lodoantipyrine autoradiography and quantitative imaging bioluminescence in consecutive tissue sections at 3, 12 and 24 hr following treatment. Digital image processing and the use of a special algorithm allowed the regional interrelationship of the 2 parameters to be quantified. Measurements revealed a great heterogeneity of blood flow and ATP between and within the tumors. A pronounced reduction of blood flow and ATP in tumors was observed after hyperthermia in comparison to untreated controls. The adjacent tissue remained mostly unaffected. However, a weakly positive relationship between the 2 parameters was obtained when variables were averaged in tumors or groups. At the microregional level, the untreated tumor tissue revealed a significant, positive correlation between nutritional blood flow and ATP concentrations. This local correlation was reduced and changed with time after treatment indicating different time courses of the parameters. Hyperthermia induced a sudden decrease in blood flow, later associated with a decline in ATP. A slight recovery of both parameters was observed 24 hr after hyperthermia. The results indicate that the metabolic status of the tumor cell is critically dependent on nutritional blood flow but also on the energy requirement of the individual tumor.