bluuebunny/crossref_metadata_2025 huggingface dataset:
Trypsinogen expression in human ovarian carcinomas
Authors: Hirahara Fumiki , Miyagi Yohei , Miyagi Etsuko , Yasumitsu Hidetaro , Koshikawa Naohiko , Nagashima Yoji , Kitamura Hitoshi , Minaguchi Hiroshi , Umeda Makoto , Miyazaki Kaoru
Published: February 2007

Increased secretion of matrix metalloproteinases and serine proteinases is well known to be associated with cancer invasion and metastasis. We aimed to elucidate the implication of trypsin, a serine proteinase and a representative digestive enzyme in invasion and metastasis of human carcinomas. Northern blot, RT‐PCR and Western blot analyses and immunohisto‐chemical studies were performed to detect and analyze trypsinogen expression in 5 ovarian carcinoma cell lines and 10 human ovarian carcinoma tissues using a DNA probe for trypsinogen I, and monoclonal and polyclonal antibodies to human trypsin I. Among the 5 ovarian carcinoma cell lines, only the MCAS (mucinous cystadenocarcinoma) cell line showed a high level of trypsinogen production and mRNA expression by Western and Northern blot analyses, respectively. However, Southern blot analysis of RT‐PCR products could detect considerable levels of trypsinogen mRNA in all ovarian cancer cell lines. In Northern analysis of ovarian cancer tissues, all advanced cancer samples showed trypsinogen gene expression. Serous cystadenocarcinomas exhibited particularly high levels of gene expression. Immunohistochemical staining also detected trypsin in ovarian carcinoma tissues. In contrast, normal ovaries and tumors with low malignant potential did not show trypsinogen expression. Our results demonstrate the extra‐pancreatic production and distribution of trypsinogen in human ovarian carcinomas.
HLA‐A2 allelic microvariants in nasopharyngeal carcinoma
Authors: Ren E. C. , Law Grace C. T. , Chan S. H.
Published: February 2007

HLA‐A2‐positive nasopharyngeal carcinoma (NPC; n = 38) and normal control (n = 51) populations were analysed by high‐resolution oligotyping to identify A2 allelic microvariants. Within the control group, A*0207 was found to be the most common allele, accounting for 50% of the A2 frequency in normals. In contrast, this allele was found to be present in only 23% of NPC cases, suggesting a protective effect. Of these 9 NPC patients with A*0207, all were associated with B46, unlike in the control group, where it can be found associated with non‐B46 antigens. Another allele, A*0201, which was thought to be protective against NPC was in fact present in 39.5% of NPC patients, more than twice the frequency in controls. These data confirm that A*0201 is not a protective allele for NPC, and other factors such as the A*0207, non‐B46 haplotype are of greater importance.
A recombinant vaccinia virus expressing human prostate‐specific antigen (PSA): Safety and immunogenicity in a non‐human primate
Authors: Hodge James W. , Schlom Jeffrey , Donohue Susan J. , Tomaszewski Joseph E. , Wheeler Clyde W. , Levine Barry S. , Gritz Linda , Panicali Dennis , Kantor Judy A.
Published: February 2007

Prostate‐specific antigen (PSA) is. a serine protease secreted by prostatic epithelial cells and is widely used as a marker for prostate cancer. The tissue specificity of PSA makes it a potential target for active specific immunotherapy, especially in prostate cancer patients who have undergone prostatectomy and in whom the only PSA‐expressing tissue in the body resides in metastatic deposits. We report here the cloning, construction and immunological consequences of immunization of rhesus monkeys with a recombinant vaccinia virus expressing human PSA (designated rV‐PSA). The prostate gland of the rhesus is structurally and functionally similar to the human prostate. While rodent and other mammalian species do not share homology with human PSA, there is 94% homology between the amino acid sequences of rhesus and human PSA. Immunization of rhesus monkeys with wild‐type vaccinia virus or rV‐PSA elicited the usual low‐grade constitutional symptoms of vaccinia virus infection. There was no evidence of any adverse effects in any immunized monkeys. A short‐lived PSA‐specific IgM antibody response was noted in all rV‐PSA immunized monkeys regardless of dose level. All monkeys receiving the 108pfu dose of rV‐PSA demonstrated PSA‐specific T‐cell responses that were maintained up to 270 days. No differences in anti‐PSA immune responses or toxicity were observed in animals that received prostatectomy prior to immunization. Our results thus demonstrate the safety and immunogenicity of rV‐PSA in a non‐human primate and have implications for potential specific immunotherapy protocols using PSA as a target.
Protein tyrosine kinase expression during the estrous cycle and carcinogenesis of the mammary gland
Authors: Andres Anne‐Catherine , Zuercher Gisela , Djonov Valentin , Flueck Martin , Ziemiecki Andrew
Published: February 2007

The susceptibility of the mammary epithelium to neoplastic transformation is linked to the exposure to estrogen during the estrous cycle. The effects of the estrous cycle on the mouse mammary gland have been investigated by analyzing <3>H‐thymidine incorporation, milk protein gene expression and DNA fragmentation. We found that the mammary epithelium undergoes limited proliferation, differentiation and apoptosis in a cycle‐dependent manner. The estrous‐responsive regulators of the mammary epithelium are unknown; however, considering the integral role of protein tyrosine kinases (PTKs) in the control of normal and malignant development, members of this family of enzymes are likely candidates for such regulatory molecules. Using a RT‐PCR‐based cloning strategy, we have undertaken a survey of PTKs expressed in the mammary gland at defined stages of development, with special emphasis on the estrous cycle. We identified 21 known and 4 novel PTKs. Their expression was analyzed throughout mammary gland development and in mammary neoplasias using a transgenic mouse model for invasive and non‐invasive carcinogenesis. Most of the identified PTKs showed highest expression during the estrous cycle and were down‐regulated during pregnancy and lactation. Deregulated expression was rarely observed in the non‐invasive mammary tumors. In contrast, 10 of 19 PTKs expressed during the estrous cycle were also over‐expressed in the invasive carcinomas, mostly involving members of the receptor family of PTKs.
Molecular heterogeneity of tumor rejection antigen/heat shock protein GP96
Authors: Feldweg Anna M. , Srivastava Pramod K.
Published: February 2007

Glycoproteins of 96 kDa (gp96) have been shown to mediate tumor‐specific immunogenicity of a number of murine sarcomas. The purity of gp96 preparations used in these studies was originally demonstrated by their homogeneity on silver‐stained gels and the observation that a single amino acid terminus was detected by micro‐sequencing. Results reported here show that gp96 preparations consist of 3–4 closely spaced bands. However, each of the bands is recognized by well‐characterized monoclonal or monospecific antibodies to gp96. We have obtained purified preparations of a slowly migrating 110‐kDa and a faster migrating 96‐kDa band of the gp96 cluster from the Meth A sarcoma and have observed both species to be immunogenic in a dose‐restricted manner. In addition to the size‐based heterogeneity, purified 96‐kDa molecules are found to contain 2 major populations, which share the same amino and carboxy termini but differ in the region recognized by an anti‐grp94 monoclonal antibody (MAb). The heterogeneity in gp96 preparations does not result from differences in glycosylation but may result from differential phosphorylation, conformation or the antigenic peptides associated with gp96. It has been suggested that the tumor‐specific immunogenicity of gp96 preparations does not derive from gp96 but from a 110‐kDa protein which co‐purifies with gp96 and is distinct from it. Our observations show instead that the presence of a number of bands in purified gp96 preparations is due to heterogeneity within gp96 molecules rather than to contamination with unrelated proteins.
Cancer incidence among filipinos in los angeles county, 1972–1991
Authors: Bernstein Leslie , Miu Angela , Monroe Kris , Henderson Brian E. , Ross Ronald K.
Published: February 2007

Although Filipinos are the second largest Asian subgroup in the United States, little is known about their patterns of cancer incidence. We have examined cancer incidence rates among Filipinos living in Los Angeles County from 1972 through 1991 and evaluated their risk of non‐localized cancer relative to non‐Hispanic whites. Although Filipinos have substantially lower cancer rates than non‐Hispanic whites in Los Angeles County, their incidence rates of liver cancer, cancer of the nasopharynx and thyroid cancer are higher. Filipino men and women had somewhat more advanced cancer at diagnosis, on average, than non‐Hispanic whites, though no statistically significant differences were observed for sites where early detection methods exist. Our data suggest that Filipino‐Americans would benefit from increased screening for cervical and breast cancer as well as efforts to interrupt transmission of hepatitis B virus to reduce liver cancer incidence.
A case‐control investigation on cancer of the ovary in Bangalore, India
Authors: Nandakumar Ambakumar , Anantha Narayanappa , Dhar Murali , Ahuja Vijaykumar , Kumar Rekha , Reddy Sreerama , Venugopal Thalagavadi , Rajanna  , Vinutha A. T. , Srinivas
Published: February 2007

Cancer of the ovary is the sixth leading cancer among females in Bangalore, and is a leading site of cancer in other population‐based cancer registries in India. A case‐control investigation was conducted utilizing the data from the population‐based cancer registry in Bangalore. In addition to the core patient information, certain other details pertaining to consumption of tobacco, reproductive and obstetric factors and those related to the practice of family planning, including the method adopted, were available with the registry, for the period 1982–1985. Identical information was also available for patients residing in the registry area who did not have cancer. Ninety‐seven cases of ovarian cancer in ever‐married women were age‐matched with 194 controls from the same area who showed no evidence of cancer. The risk of ovarian cancer was not influenced by tobacco habits, alcohol consumption, diet or the various reproductive factors. However, tubectomy as a method of family planning appeared to reduce the risk of development of ovarian cancer. This reduction in risk was not influenced by parity or age of the woman at the time of birth of the first child.
Life‐long over‐expression of ornithine decarboxylase (ODC) gene in transgenic mice does not lead to generally enhanced tumorigenesis or neuronal degeneration
Authors: Alhonen Leena , Halmekytö Maria , Kosma Veli‐Matti , Wahlfors Jarmo , Kauppinen Risto , Jänne Juhani
Published: February 2007

Ornithine decarboxylase, the rate‐controlling enzyme of the biosynthetic pathway of the polyamines, is one of the most inducible mammalian enzymes showing many of the features common to the oncoproteins. Ornithine decarboxylase activity is likewise strongly induced in response to various neurotoxic stimuli, and the enhanced enzyme activity is believed to be causally related to neuronal damage. We have generated several transgenic mouse lines over‐expressing human ornithine decarboxylase gene. We have now subjected the animals to a long‐term survival study in order to assess whether constitutively over‐expressed ornithine decarboxylase would predispose them to enhanced tumorigenesis and neuronal degeneration. When the transgenic animals were 2 years old, their tissue ornithine decarboxylase activity was 20‐ to 50‐fold that in their syngenic littermates. Macroscopic and microscopic examination of organs revealed no differences between syngenic and transgenic animals as regards spontaneous tumor incidence or age‐related changes in brain.
Heterogeneity in production, secretion and glycosylation of MUC1 epithelial mucin by primary cultures of ovarian carcinoma
Authors: McGuckin Michael A. , Hurst Terence G. , Ward Bruce G.
Published: February 2007

The MUCI mucin produced by many adenocarcinomas has functions that may be of biological significance and is of importance clinically as a serum tumour marker and as a candidate target for immunotherapy. Previous studies of MUCI production by ovarian cancers have been limited to immunohis‐tochemical studies of tumour specimens and in vitro studies using cell lines. In this study the biosynthesis, secretion and glycosylation of MUCI were studied in primary cultures of tumour cells obtained from 35 patients with stage 3 ovarian cancer. Although 34 of the 35 tumours produced MUCI in vitro, the concentrations of intracellular and secreted MUCI, as measured by an ELISA using core protein‐reactive antibodies, varied over a wide range. In addition, the amount of secreted MUCI as a proportion of the intracellular concentration varied between tumours. Pulse/chase amino acid labelling studies of MUCI biosynthesis also demonstrated variation in secretion rates. Multivariate regression analysis showed that of the variables tumour size, histological type, grade, ploidy status and intracellular and secreted MUCI concentrations in vitro, only mucin secretion rate was significantly associated with serum mucin concentrations (p < 0.001). Culture of tumour cells for 4 days in the presence or absence of a competitive inhibitor of O‐glycosylation, BAG, showed that the degree of glycosylation of MUCI varied between tumours and that under‐glycosylation was not correlated with production or secretion rates. Our study has shown heterogeneity in the production, secretion and glycosylation of MUCI and a strong correlation between the secretion rate in vitro and the concentration in the serum of patients.
Human esophageal carcinomas frequently express the tumor‐rejection antigens of MAGE genes
Authors: Inoue Hiroshi , Mori Masaki , Li Jian , Mimori Koshi , Honda Masayuki , Nakashima Hideaki , Mafune Ken‐Ichi , Tanaka Youichi , Akiyoshi Tsuyoshi
Published: February 2007

Thde human genes MAGE‐1 and ‐3 encode melanoma peptide antigens that are recognized bhd autologous cytotoxic T lymphocytes. Tumors expressing MAGE genes are potential targets for cancer immunotherapy, because MAGE genes are expressed only in tumor tissue and not in any normal tissue except testis and placenta. However, little is known about MAGE gene expression in human esophageal carcinoma. The purpose of this study was therefore to analyze MAGE gene status in human esophageal carcinoma. We studied the expression status of these genes in 42 surgical samples and in 12 cell lines of human esophageal carcinoma using the reverse transcription polymerase chain reaction (RT‐PCR). Various clinicopathological factors were also analyzed. No MAGE gene expression was seen in any of the 42 normal esophageal tissue specimens. In contrast, tumor tissue expressed MAGE‐1, ‐2, and ‐3 in 26, 18 and 24 specimens, respectively. Thirty‐three of 42 tumors expressed at least one MAGE gene. Significant clinicopathologic differences between the tumors were not observed, regardless of the presence or absence of MAGE gene expression. In cell lines, MAGE‐1, ‐2, and ‐3 gene expression was recognized in 5, 4 and 4 cell lines, respectively. This study demonstrates that MAGE genes are frequently expressed in clinical samples as well as in cell lines of esophageal carcinoma. The identification of MAGE genes, therefore, may open up a new modality of treatment, namely specific immunotherapy, for patients with esophageal carcinoma.
Mutant androgen receptors in prostatic tumors distinguish between amino‐acid‐sequence requirements for transactivation and ligand binding
Authors: Peterziel Heike , Culig Zoran , Stober Jutta , Hobisch Alfred , Radmayr Christian , Bartsch Georg , Klocker Helmut , Cato Andrew C. B.
Published: February 2007

Mutant androgen receptors are thought to contribute to hormone resistance in prostate carcinoma. The part they play in this process, however, is ill‐defined. Here we report on transactivation by 2 mutant androgen receptors from prostatic tumors with single amino‐acid exchanges in their hormone‐binding domains. These exchanges enhance the transactivation property of the receptors, particularly to androsterone and androstanediol, 2 metabolized derivatives of testosterone present in the prostate. Additionally, they enhance the transactivation potential of the mutant receptors to hydroxyflutamide, an anti‐androgen frequently used in hormone ablation therapy. The increased transactivation by the mutant receptors did not result from altered affinity of the receptors to the inducing ligands nor from measurable changes in conformation of the liganded receptors. Thus the single amino‐acid exchanges identify differences in amino‐acid‐sequence requirements for transactivation and ligand binding in the hormone‐binding domain of the androgen receptor. These results provide new insights into ligand‐dependent transactivation, and form a framework for the search for effective antagonists to be used in prostate‐cancer therapy.
gelatinase A/TIMP‐2 imbalance in lymph‐node‐positive breast carcinomas, as measured by RT‐PCR
Authors: Onisto Maurizio , Riccio Maria Pia , Scannapieco Paolo , Caenazzo Cristina , Griggio Luciano , Spina Michele , Stetler‐Stevenson William G. , Garbisa Spiridione
Published: February 2007

Over‐production of gelatinase A (MMP‐2) or under‐production of its inhibitor (TIMP‐2) may result in the matrix degradation crucial for metastasis, and early evaluation of their expression in primary tumor would offer important prognostic informations. RT‐PCR amplicons of MMP‐2 and TIMP‐2 mRNA from tissue biopsies of 13 breast carcinomas and one fibrocystic mastopathy were quantitated. In comparison with their normal‐tissue counterparts, their expression trends were not uniform: in some cases MMP‐2 increased in the tumor without changes in TIMP‐2, in others TIMP‐2 expression also increased, although to a lesser extent than MMP‐2; only in 2 cases was it slightly lower in the tumor tissue. Nevertheless, clearer insights were gained from the comparison of the ratio (R) between MMP‐2tumor/normal and TIMP‐2tumor/norma: as in the fibrocystic mastopathy, the R in carcinomas without lymph‐node involvement (LN−) was usually lower than 1 in most cases. In contrast, in 5 out of 6 patients with lymph‐node metastasis (LN+), the ratio ranged between 2 and 4. While the R magnitude was not related to the frequency of positive lymph nodes out of the total analyzed, nor to relapse status at follow‐up (all relapse‐free), the clear‐cut difference between the LN− and LN+ groups was statistically significant. Results suggest that evaluation of MMP‐2/TIMP‐2 mRNA balance may constitute an early prognostic approach, which may also be more reliable concerning cancer aggressiveness as compared with the MMP‐2 alone, and that boosting TIMP‐2 expression may be a therapeutic strategy to prevent metastasis. © 1995 Wiley‐Liss, Inc.
Dendritic antigen‐presenting cells from the peripheral blood of renal‐cell‐carcinoma patients
Authors: Radmayr Christian , Böck Günther , Hobisch Alfred , Klocker Helmut , Bartsch Georg , Thurnher Martin
Published: February 2007

Dendritic cells are considered to be the initiators of immune responses, including those directed against tumors. Clinical research on dendritic cells was long hampered by the limited availability of these cells. The recent identification of cytokine combinations that mobilize dendritic cells with potent antigen‐presenting cell function from peripheral blood represented a major progress. We show in this study that substantial numbers of dendritic cells can be obtained from the peripheral blood of patients with renal‐cell carcinoma. The procedure requires a relatively small blood sample (40 ml) and avoids both priming of the patient with granulocyte‐colony stimulating factor and leukapheresis. Approximately 2 to 8 million cells with the characteristics of dendritic cells could be obtained: phase‐contrast microscopy revealed the typical cytoplasmic processes or veils; phenotypic analysis confirmed expression of dendritic‐cell‐associated molecules, including MHC class II, CDIa, CD4, ICAM‐1 (CD54), LFA‐3 (CDS8), B7‐1 (CD80) and B7‐2 (CD86), and absence of T‐cell, B‐cell and monocyte markers; in addition, these cells rapidly attached to and migrated on collagen‐type‐1‐coated surfaces. Interestingly, attachment was accompanied by acquisition of the CD14 antigen; functionally, cultured dendritic cells proved to be very potent co‐stimulators of the phytohemag‐glutinin‐induced proliferation of autologous tumor‐infiltrating lymphocytes. The reproducible growth of functional dendritic cells from cancer patients is encouraging for the design of immunotherapy protocols. © 1995 Wiley‐Liss, Inc.
Somatic mutations of von Hippel‐Lindau (VHL) tumor‐suppressor gene in european kidney cancers
Authors: Bailly Maryse , Bain Christine , Favrot Marie‐C. , Ozturk Mehmet
Published: February 2007

Somatic mutations of von Hippel Lindau (VHL) tumor‐suppressor gene have been identified in kidney cancers from North America and Japan. We studied VHL gene mutation in 31 kidney tumors from France. Of these tumors, 45% (14/31) displayed mutations, 60% of which occurred at A:T base pairs. The frequency and the spectrum of mutations identified in kidney tumors from Europe were similar to those reported for tumors from other geographical locations. © 1995 Wiley‐Liss, Inc.
Neural and mesenchymal differentiations in Ewing's sarcoma cell lines. Morphological, immunophenotypic, molecular biological and cytogenetic evidence
Authors: Pagani A. , Fischer‐Colbrie R. , Eder U. , Pellin A. , Llombart‐Bosch A. , Bussolati G.
Published: February 2007

Three established Ewing's sarcoma (ES) cell lines (TC106, 6647, A4573), grown both in vitro and as xenograft tumors, were analyzed. In all 3 lines and tumors, the ES characteristic reciprocal translocation (11;22), as well as the presence of the ES‐associated p30/32M1C2 antigen, were documented. However, these cell lines showed discrepancies in their neural and mesenchymal differentiation. The TC106 line was characterized by expression of the neuroendocrine marker secretogranin II (SgII) which was detectable by Northern blot and by radioimmunological detection (RIA) in the culture medium of secretoneurin, a proteolytic product of SgII. In contrast, TC106 cells were immunohistochemically and radioimmunologically secretoneurin‐negative. Rapid cellular secretion of the peptide is probably the explanation of such a discrepancy. The 6647 and A4573 cell lines were SgII/secretoneurin‐negative and the former presented morphological and immunocytochemical evidence of mesenchymal differentiation. In fact, the 6647 xenograft tumor showed osteosarcomatous‐type morphological features and the neoplastic cells were immunocytochemically positive for osteonectin and osteocalcin antigens. Expression of the CgA gene, which is typical of neuroblastomas, was absent in the Ewing's sarcoma cell lines investigated. Our findings provide experimental evidence that Ewing's sarcoma is a heterogeneous tumor which may show either neuroectodermal or mesenchymal differentiation. SgII/secretoneurin analysis by hybridization and RIA procedures is a reliable approach to the identification of ES with neuroendocrine differentiation. © 1995 Wiley‐Liss, Inc.
Metastatic behavior of human tumors in congenitally athymic nude mice: Intrinsic properties of the tumor cells and host immune reactivity
Authors: Rofstad Einar K.
Published: February 2007

Congenitally athymic nude (nu/nu) mice have been used extensively as hosts to study the metastatic properties of xenografted human tumors. It has not been fully explored to what extent the metastatic behavior of human tumor xenografts reflects the intrinsic properties of the tumor cells and to what extent it is influenced artificially by the host. The purpose of the work reported here was to evaluate the potential usefullness of athymic mice for qualitative and quantitative assessment of the intrinsic metastatic properties of human tumor cells. Four human melanoma cell lines (A‐07, D‐12, R‐18, U‐25) were included in the study. Quantitative intradermal and intracranial transplantation assays were used to determine the tumorigenicity and immunogenicity of the cell lines. The metastatic behavior of the cell lines was studied by inoculating cells intra‐arterially or intravenously (artificial metastases) or intra‐dermally (spontaneous metastases). The influence of the host on the incidence of metastases was assessed by inoculating cells intravenously in immunologically modified athymic mice. Both the intradermal and intracranial tumorigenicity differed substantially between the cell lines. The host immune reactivity against heterotransplantation was significantly lower for D‐12 than for A‐07, R‐18 and U‐25 cells. The incidence of metastases was influenced significantly by the host immune reactivity. The cell lines showed organ‐specific metastatic patterns in athymic mice. The organs of preference were lungs for A‐07 and D‐12 cells, lymph nodes for R‐18 cells, and brain for U‐25 cells. The organ‐specific metastatic pattern in athymic mice reflected the pattern of distant metastases in the donor patients. In addition, all cell lines gave rise to metastases in abdominal organ sites, but without organ specificity. The incidence of abdominal metastases was influenced significantly by the tumorigenicity of the cell lines. In conclusion, athymic mice appear to be excellent hosts for assessment of the organ specificity in the metastatic behavior of human tumors. The metastasis frequency of human tumors in athymic mice, however, might be a misleading quantitative measure of the intrinsic metastatic propensity of the tumor cells, owing to the cell‐line‐dependent host immune reactivity. © 1995 Wiley‐Liss, Inc. © 1995 Wiley‐Liss, Inc.
Family history of cancer is a risk factor for squamous cell carcinoma of the head and neck in Brazil: A case‐control study
Authors: Foulkes William D. , Brunet Jean‐Sebastien , Kowalski Luis P. , Narod Steven A. , Franco Eduardo L.
Published: February 2007

To determine the role of familial factors in head and neck cancer, we analysed data from a hospital‐based case‐control study of squamous cell carcinoma of the head and neck in Brazil. There were 754 cases of squamous cell carcinoma of the head and neck (SCCHN) and 1,507 age‐ and gender‐matched hospital‐based controls with non‐malignant diseases. Subjects provided information on the occurrence of cancer in first‐degree relatives, as well as about other risk factors, including tobacco and alcohol consumption. Relative risks (RRs) were estimated for developing mouth, pharynx and larynx cancer when cancers in relatives were observed. RRs were adjusted for age, sex, city of admission and alcohol and tobacco consumption. The RR for developing SCCHN if a first‐degree relative had cancer at any site was significantly elevated at 1.97. The RR was 3.65 (95% Cl: 1.97–6.76) if the relative had head and neck cancer. Significantly elevated risks for developing head and neck cancer were associated with siblings with head and neck cancer (RR = 8.57) and, to a lesser extent, with fathers with head and neck cancer (RR = 2.49). There was no significantly increased risk associated with mothers with head and neck cancer, but these tumours were rare among mothers. Our data show that familial, possibly genetic, factors are important in the aetiology of head and neck cancer. © 1995 Wiley‐Liss, Inc.
Clinical usefulness of free PSA fraction as an indicator of prostate cancer
Authors: Filella Xavier , Alcover Juan , Molina Rafael , Giménez Núria , Rodríguez Alfredo , Jo Judith , Carretero Pablo , Ballesta Antonio Manuel
Published: February 2007

The usefulness of the fPSA fraction in the differential diagnosis of benign prostate hyperplasia (BPH) and prostate cancer was evaluated, with the aim of improving the diagnostic efficacy of PSA. Serum PSA and fPSA determinations were performed by an enzymoimmunoassay technique on an ES‐300 system. fPSA constitutes a minor fraction both in normal subjects and in patients with prostate disease, being significantly lower in patients with untreated prostate cancer than in patients with BPH. Likewise, the authors have observed that the sensitivity of the fPSA/PSA ratio has an inversely proportional relationship with the stage of the disease. The results obtained in patients with PSA levels between 4 and 20 μg/l are also of note. In this series of patients, the efficacy of the fPSA/PSA ratio was higher than that of PSA, showing a sensitivity of 44% and a specificity of 95% at the cut‐off of 0.08. © 1995 Wiley‐Liss, Inc.
Adhesion molecules involved in the binding of murine myeloma cells to bone marrow stromal elements
Authors: Okada Tomoko , Hawley Robert G.
Published: February 2007

In previous work, we reported the development of the B9/BM1 syngeneic murine bone marrow metastasis model. Interleukin (IL)‐6–dependent, IL‐1–producing B9/BM1 cells, which preferentially home to and colonise the vertebral and femoral marrow after i.v. injection, exhibit striking similarity in cell surface phenotype to human myeloma cells, especially the expression of 3 adhesion molecules, CD44, VLA‐4 and ICAM‐I. Because the haematopoietic microenvironment consists of different cell types, such as endothelial cells, fibroblasts, adipo‐cytes and macrophages, we investigated the functional significance of these adhesion molecules in heterotypic binding assays between B9/BM1 cells and a newly established bone marrow‐derived endothelial cell line (BMEC), a fibroblastoid pre‐adipocyte cell line (BMS2.2) and primary bone marrow‐derived macrophages. B9/BM1 cells adhered well to all stromal elements: a combination of monoclonal antibodies (MAbs) against CD44 and VLA‐4 significantly inhibited the adherence of B9/ BM1 cells to BMEC and BMS2.2 cells, whereas binding of B9/BM1 cells to macrophages was partially blocked with an anti‐ICAM‐I MAb. Our results implicate multiple recognition mechanisms, including those involving CD44, VLA‐4 and ICAM‐I, in the retention of B9/BM1 cells in the bone marrow. © 1995 Wiley‐Liss, Inc.
Synthesis, maturation and extracellular release of procathepsin D as influenced by cell proliferation or transformation
Authors: Isidoro Ciro , Demoz Marina , De Stefanis Daniela , Baccino Francesco M. , Bonelli Gabriella
Published: February 2007

The relationship between cell growth and intra‐ and extracellular accumulation of cathepsin D (CD), a lysosomal endopepti‐dase involved in cell protein breakdown, was examined in cultures of normal and transformed BALB/c mouse 3T3 fibro‐blasts grown at various cell densities. In crowded cultures of normal 3T3 cells (doubling time, Td, 53 hr) intracellular CD activity was 2–fold higher than in sparse, rapidly‐growing (Td, 27 hr) cultures. In uncrowded (Td, 18 hr) and crowded (Td, 32 hr) cultures of benzo[a]pyrene‐transformed cells intracellular CD levels were one third and two thirds, respectively, of those measured in hyperconfluent 3T3 cultures. Regardless of cell density, SV‐40–virus‐transformed cells (Td, 12 hr) contained one third of CD levels found in hyperconfluent 3T3 cells. Both transformed cell lines released into the medium a higher proportion of CD, compared with their untransformed counterpart, yet the amount secreted was not sufficient to account for the reduced intracellular level of the enzyme. Serum withdrawal induced a marked increase of both intra‐ and extracellular levels of CD activity. In both normal and virally or chemically transformed 3T3 cells CD comprised a precursor (52 kDa) and processed mature polypeptides; the latter were mostly represented by a 48–kDa peptide, but a minor part was in a double‐chain form (31 and 16 kDa respectively). The proportion of mature enzyme vs. precursor was much higher in confluent, slowly‐growing cells than in fast‐growing cells, whether normal or transformed. In the latter, conversion of mature 48–kDa peptide into the double‐chain form occurred more efficiently. © 1995 Wiley‐Liss, Inc.
Analysis and clinical implications of k‐ras gene mutations and infection with human papillomavirus types 16 and 18 in primary adenocarcinoma of the uterine cervix
Authors: Tenti Patrizia , Romagnoli Solange , Silini Enrico , Pellegata Natalia S. , Zappatore Rita , Spinillo Arsenio , Zara Carlo , Ranzani Guglielmina N. , Carnevali Luciano
Published: February 2007

Experimental models indicate that activated ras genes and HPV oncogenic sequences may cooperate in inducing a completely transformed phenotype in epithelial cells. We searched for K‐ras gene mutations and HPV type‐16 and ‐18 sequences in 67 primary adenocarcinomas of the uterine cervix by analyzing DNAs from formalin‐fixed, paraffin‐embedded tissue samples. Target sequences were amplified by PCR and analyzed by denaturing gradient gel electrophoresis (DGGE) and sequencing for the detection of K‐ras gene mutations and by Southern blotting for the detection of HPV infection. We found 16 mutations in 15 cases; 14 were at codon 12 and 2 at codon 13; 11 were base transitions and 5 were transversions. Mutations were more frequent in mucin‐secreting than in non‐mucinous tumors. HPV oncogenic sequences were detected in 58 cases with no significant difference between K‐ras‐mutated and wild‐type tumors. HPV oncogenic sequences were also more frequent in mucin‐secreting than in non‐mucinous tumors. Both molecular events were present simultaneously in 13 out of 58 cases, all of which had histologically grade‐2 and grade‐3 tumors. Clinico‐pathological parameters of the disease and the overall survival, however, were independent of K‐ras mutations and of HPV‐16 and ‐18 infection, as shown by univariate and multivariate analysis. In contrast, stage of disease, lymph‐node metastases, deep infiltration, clear‐cell histology and low grade of differentiation were risk factors for tumor‐related death. © 1995 Wiley‐Liss, Inc.
Interphase cytogenetic analysis of erbB2 and topollα co‐amplification in invasive breast cancer and polysomy of chromosome 17 in ductal carcinoma in situ
Authors: Murphy Dermot S. , McHardy Peter , Coutts Jacqueline , Mallon Elizabeth A. , David George W. , Kaye Stanley B. , Brown Robert , Nicol Keith W.
Published: February 2007

Breast cancer is a genetically complex disease. Fluorescence in situ hybridisation can be used to analyse the genetics of breast‐cancer progression by interphase cytogenetics. We have analysed the histological distribution of erbB2 and topollα co‐amplification in paraffin sections of invasive breast cancer and show that the co‐amplified loci share the same histological distribution in the tumour and have a similar nuclear distribution within individual nuclei. Regions of the tumours without amplification are easily recognized and tumours with erbB2 and topollα co‐amplification can be distinguished from those with erbB2 amplification alone. In addition, FISH was used to show polysomy of chromosome 17 in non‐invasive ductal carcinoma in situ of the breast and erbB2 amplification in both the invasive and non‐invasive components of a breast cancer biopsy. This report of an interphase cytogenetic analysis of non‐invasive breast carcinoma in situ demonstrates the usefulness of FISH for the genetic study of breast cancer progression. © 1995 Wiley‐Liss, Inc.
Expression of galectin‐1 in normal human thyroid gland and in differentiated and poorly differentiated thyroid tumors
Authors: Chiariotti Lorenzo , Berlingieri Maria Teresa , Battaglia Caterina , Benvenuto Giovanna , Martelli Maria Luisa , Salvatore Paola , Chiappettaxy Gennaro , Bruni Carmelo Bruno , Fusco Alfredo
Published: February 2007

We previously reported that galectin‐l gene expression increases up to 100‐fold in oncogene‐transformed rat thyroid cells compared with their normal counterparts and that the relative mRNA levels correlate with the degree of malignancy. In the present study we investigated whether galectin‐l is differentially expressed in human thyroid neoplasms, which range from well‐differentiated tumors to undifferentiated ana‐plastic carcinomas. We analyzed 74 human thyroid specimens of neoplastic, hyperproliferative and normal tissues and several tumor cell lines. Galectin‐l mRNA and protein levels were higher in 6 thyroid carcinoma‐derived cell lines than in normal thyroid primary cultures and adenoma cells. Galectin‐l mRNA levels increased in 28/40 papillary carcinomas and in 6/7 anaplastic carcinomas compared with normal or hyperplastic thyroid. Conversely, galectin‐l expression was unaffected in follicular carcinomas and benign adenomas. Immunohistochemi‐cal analysis of normal thyroid and papillary carcinoma sections revealed a higher content of galectin‐l protein in neoplastic follicular cells than in normal cells. © 1995 Wiley‐Liss, Inc.
p53 antibodies in the sera of lung cancer patients: Comparison with p53 mutation in the tumour tissue
Authors: Wild Christopher Paul , Ridanpaa Maaret , Anttila Sisko , Lubin Richard , Soussi Thierry , Husgafvel‐Pursiainen Kirsti , Vainio Harri
Published: February 2007

This study examined the sensitivity and specificity of serum auto‐antibodies to p53 protein as a non‐invasive marker of p53 genetic alterations or protein accumulation in lung cancer cases. A sensitive ELISA to detect serum p53 antibodies was developed and used to examine sera from 186 patients undergoing pulmonary surgery for a suspected lung cancer. Target antigens in ELISA were wild‐type p53 protein and 5 peptides covering the N‐ and C‐terminal parts of the protein. Sixteen sera were positive for serum p53 antibodies in both ELISAs and all were among the 136 patients with confirmed primary lung carcinoma. Of 50 patients with other pulmonary diseases, none had p53 antibodies. In 92 cancer patients exons 5 to 8 of the p53 gene were examined for mutations by denaturing gradient gel electrophoresis and direct sequencing of PCR products. Forty‐seven tumours had a p53 mutation and 7 (15.2%) of these were positive for p53 antibodies. Two patients had serum antibodies but no detectable mutation in exons 5 to 8. Frequencies of p53 mutations and serum antibodies were higher in squamous cell carcinoma patients than in adenocarcinoma. Accumulation of p53 protein in tumour tissue was observed in 32 patients, but only 5 were positive for p53 antibodies. In conclusion, serum p53 antibodies were detected only in a proportion of lung cancer cases, but the majority were specifically associated with a detectable p53 mutation in the tumour. © 1995 Wiley‐Liss, Inc.
Can a test for E6/E7 transcripts of human papillomavirus type 16 serve as a diagnostic tool for the detection of micrometastasis in cervical cancer?
Authors: Czeglédy Judit , Iosif Constantin , Hansson Bengt Göran , Evander Magnus , Gergely Lajos , Wadell Göran
Published: February 2007

Tissue from 11 cases of cervical cancer positive for human papillomavirus (HPV) type 16 DNA and 69 pelvic lymph nodes from the same patients were examined for HPV 16 DNA and mRNA from the E6/E7 genes. Five of the tumors were squamous, 3 adeno‐ and 3 adenosquamous carcinoma. From the primary tumors and the extirpated lymph nodes DNA and RNA or mRNA was subjected to PCR and RT‐PCR. Three transcription profiles (only E6*1, E6*1 and E6*11 or full‐length E6‐E7 plus both of the spliced transcripts) were found in all of the 11 HPV 16 DNA‐positive primary tumors. From the total of 69 lymph nodes analyzed 28 were positive for mRNA. HPV 16 DNA was found in 7 additional samples. Cytokeratin was found in 19 of these lymph nodes, indicating epithelial origin of tumor cells. Only 1 patient had 2 metastases evidenced by histology. These were both positive for HPV DNA and mRNA. The finding of HPV DNA, mRNA and cytokeratin in lymph nodes of patients with cervical cancer should be an indication of lymphogenically driven micrometastases of the tumor. The HPV mRNA assay should offer higher specificity than the DNA test since mRNA can be found in live cells only, while HPV DNA also can originate from dead cell material sequestered in the lymph nodes. © 1995 Wiley‐Liss, Inc.
Analysis of the gap‐related domain of the neurofibromatosis type 1 (NF1) gene in childhood brain tumors
Authors: Scheurlen Wolfram G. , Senf Leonore
Published: February 2007

rasThe neurofibromatosis type‐1 (NF1) gene contains a 360‐bp region with significant homology to the catalytic domain of mammalian GTPase‐activating protein. This particular GAP‐related domain of the NF1 gene (NF1‐GRD) stimulates ras GTPase and inactivates ras protein p21. Therefore, it has been suggested that the NF1 gene represents another tumor‐suppressor gene. In the search for molecular markers of possible diagnostic relevance, childhood brain‐tumor specimens of different histologic diagnoses were tested for mutations of the so‐called FLR‐exon within the NF1 ‐GRD. This part of the NF1‐GRD has been shown to be most crucial for the GAP‐like function. Using a highly sensitive PCR‐SSCP technique, we tested 51 tumor specimens were tested, but found no mutations. We conclude that inactivation of this putative tumor‐suppressor gene by mutations does not play a significant role in tumorigenesis of childhood brain tumors. Next, we compared the splice variants of the NF1‐GRD in 33 brain tumors and 8 extraneural embryonal tumors. Primitive neuroectodermal tumors (PNET) (n = 10) and one intracranial teratoma were the only tumors that predominantly expressed a splice pattern that can be observed in the immature developing brain. In contrast to other embryonal neuronal tumors, this NF1‐GRD splicing pattern could not be modified in a newly established medullo‐blastoma cell line by retinoic acid treatment. Since this particular splice variant suppresses p21ras more effectively than other NF1‐GRD transcripts, its predominant expression may interfere with the physiological signal transduction of p21ras during differentiation of neurons. There may be a neurofibromin‐induced and p21ras‐mediated differentiation pathway of neuronal stem cells that is blocked in PNET. Such an arrest of a p21ras‐dependent differentiation pathway may explain the persistence of primitive pluripotent neuronal cells in PNET. © 1995 Wiley‐Liss, Inc.
Expression of α3β1 integrin receptor and its ligands in human lung tumors
Authors: Bartolazzi Armando , Cerboni Cristina , Flamini Giovanna , Bigotti Aldo , Lauriola Libero , Natali Pier Giorgio
Published: February 2007

Increasing experimental evidence demonstrates that malignant transformation is associated with changes in the repertoire of expression of the integrin family of molecules, which mediate cell‐matrix and cell‐cell interactions. We have analyzed immuno‐histochemically and immunochemically the expression of VLA‐3 integrin and its known ligands, namely, laminin (LM), fibronectin (FN), collagen type IV (Coll IV), nicein (NIC), and entactin/nidogen (ENT), in lung tumors of various histological types. α3β1 was detectable in normal bronchial epithelium and along basement membranes of alveolar walls. In non‐small cell lung carcinomas (NSCLC) the integrin was expressed in 82% of the cases, independently of histological type and degree of differentiation of the tumors. On the other hand, only 13% of the small cell lung carcinomas (SCLC) displayed a weak and heterogeneous distribution of the α3β1 complex. Our findings were confirmed immunochemically using long‐term tumor cell lines. While the expression of both α3β1 and ligands LM, FN, Coll IV, and Ent correlated in NSCLC with the presence of basement membranes, FN was the only ligand detectable in the stroma of SCLCs. A selective loss of nicein in basement membranes was demonstrated in NSCLC indicating an impairment of expression of this glycoprotein following malignant transformation. © 1995 Wiley‐Liss, Inc.
Microsatellite instability and pathological aspects of breast cancer
Authors: Contegiacomo Alma , Palmirotta Raffaele , De Marchis Laura , Pizzi Claudia , Mastranzo Paolo , Delrio Paolo , Petrella Giuseppe , Figliolini Marino , Bianco Angelo Raffaele , Frati Luigi , Cama Alessandro , Mariani‐Costantini Renato
Published: February 2007

Genomic instability plays a key role in hereditary non‐polyposis colorectal cancer and in a significant sub‐set of non‐hereditary colorectal tumors. Recent evidence suggests that microsatellite instability also occurs in various sub‐sets of common, non‐hereditary forms of extra‐colorectal carcinoma. To investigate the role of microsatellite instability in breast cancer, and to correlate this type of alteration with clinico‐pathological characteristics, including tumor proliferative activity, we analyzed the status of 8 different microsatellite loci in 28 cases of primary mammary carcinoma. For this purpose, microsatellite banding patterns were compared in paired breast‐cancer/peripheral‐blood DNA samples. Microsatellite instability was observed in 6/28 (21%) of the cases. Four of the 6 tumors had low proliferative activity, one had high proliferative activity, and in one case proliferative activity values were not available. All chromosomal loci investigated demonstrated microsatellite instability in one or more representative tumors of the series. Shifts in length larger than 2 bp were the most frequent change. Microsatellite instability significantly correlated with the lobular histotype, and with lymph‐node involvement. A trend was also observed associating microsatellite instability and large tumor size. © 1995 Wiley‐Liss, Inc.
Microsatellite instability in japanese esophageal carcinoma
Authors: Nakashima Hideaki , Mori Masaki , Mimori Koshi , Inoue Hiroshi , Shibuta Kenji , Baba Kinva , Mafune Ken‐Ichi , Akiyoshi Tsuyoshi
Published: February 2007

Recent studies have shown that microsatellite instability (MSI) may play an important role in the development of various types of cancer. However, there have been only 2 reports describing MSI in esophageal carcinoma and the clinicopathologic significance of MSI in this malignancy has not yet been clarified. To better elucidate the role of genetic instability in the development of esophageal carcinoma, we investigated the presence of MSI in 32 cases of esophageal cancer using paired samples of fresh frozen tumor and normal tissue by a method based on the polymerase chain reaction. MSI was defined as occurring in tumors which showed altered banding patterns at one or more microsatellite loci. The incidence of MSI in esophageal carcinoma was 6 out of 32 patients. MSI was observed more frequently in cases with small‐cell carcinoma (2 out of 2) than in cases with squamous‐cell carcinoma (4 out of 29). No cases with adenocarcinoma or Barrett's metaplasia were included in our series. No significant correlations between MSI and other clinicopathologic parameters were observed. The present study suggests that (I) some Japanese esophageal carcinomas certainly correlate with DNA replication error, and (2) MSI may be more frequent in small‐cell carcinoma of the esophagus than in squamous‐cell carcinoma of the esophagus. © 1995 Wiley‐Liss, Inc.
Characterization of monoclonal antibodies against stromelysin‐3 and their use to evaluate stromelysin‐3 levels in breast carcinoma by semi‐quantitative immunohistochemistry
Authors: Santavicca Maria , Noel Agnès , Chenard Marie‐Pierre , Lutz Yves , Stoll Isabelle , Segain Jean‐Pierre , Rouyer Nicolas , Rio Marie‐Christine , Wolf Catherine , Bellocq Jean‐Pierre , Basset Paul
Published: February 2007

Stromelysin‐3 (ST3) is a matrix metalloproteinase which is expressed in fibroblastic cells of most human invasive carcinomas and represents a potential new prognostic indicator. Expression of recombinant ST3 forms in Escherichia coli from cDNA constructs indicated that high levels of expression were achieved when the ST3 pro‐domain was deleted. The putative mature form of ST3 thus produced and recovered from bacterial inclusion bodies was used to prepare monoclonal antibodies (MAbs) against ST3 by immunization of BALB/C mice. Ten hybridomas producing MAbs against ST3 were obtained and analyzed for their ability to detect endogenous ST3 in breast cancer and in conditioned media from human fibroblasts. One of these MAbs (5ST‐4A9) was found to be suitable for the routine detection of ST3 on breast cancer tissue sections, thus opening the possibility to evaluate ST3 prognostic value in breast cancer using semi‐quantitative immunohistochemistry. © 1995 Wiley‐Liss, Inc.