bluuebunny/crossref_metadata_2025 huggingface dataset:
Regulation of integrin‐mediated adhesion to laminin and collagen in human melanocytes and in non‐metastatic and highly metastatic human melanoma cells
Authors: Danen Erik H. J. , Van Muijen Goos N. P. , Jansen Kees F. J. , Ruiter Dirk J. , Van De Wiel‐van Kemenade Elly , Figdor Carl G.
Published: February 2007

We compared integrin‐mediated adhesion to extracellular matrix (ECM) components of cultured human melanocytes and 6 human melanoma cell lines with different metastatic capacities in nude mice. Cultured melanocytes and most melanoma cell lines adhered strongly to fibronectin (FN), whereas only highly metastatic cell lines adhered to laminin (LM), collagen type I (COI) and type IV (COIV). Adhesion to LM and CO could be blocked by anti‐α6 and anti‐α2 monoclonal antibodies (MAbs) respectively. This observation is consistent with the finding that expression of LM receptor α6β1 and LM/CO receptor α2β1 was low on melanocytes and non‐ or poorly metastatic cell lines, whereas these integrins were strongly expressed on highly metastatic cell lines. In addition, immunoprecipitation from [35S]‐methionine‐labeled cells demonstrated increased synthesis of α6, α2 and β1 in highly metastatic cell lines and immunohis‐tochemistry showed expression of α6β1 and α2β1 only in xenograft lesions from highly metastatic cell lines. Furthermore, the observation that adhesion of melanocytes and non‐ or poorly metastatic cell lines could be stimulated with anti β1 MAbs demonstrates that these receptors, on these cells, are expressed in an inactive state. Our results suggest that α2β1 and α6β1 play a role in human melanoma metastasis in nude mice and demonstrate that interactions of these integrins with their ligands can be regulated at the level of surface expression and activation state of the receptor.
Expression of the MuLV‐tumor‐associated antigen is restricted to MuLV‐transformed cells
Authors: Flyer David C. , Sucheski Brian
Published: February 2007

Immunization of mice with Moloney murine leukemia virus (MoMuLV) induces the generation of a population of CTL which recognizes a non‐viral, tumor‐associated antigen (TAA) expressed on MuLV‐induced tumors. To determine whether this TAA could be used as a pre‐leukemic or leukemic cell marker, CTL clones directed against Moloney viral and TAA antigen were used to analyze viral and TAA antigen expression on chronically infected and leukemic lymphoid cells obtained from mice inoculated neonatally with MoMuLV. Although both sets of cells could be recognized and lysed by viral antigen specific CTL, they are not recognized by TAA‐specific CTL. Only after transformed cell lines were established from leukemic spleen cells could susceptibility to TAA‐specific CTL be observed. Thus, the appearance of the MoMuLV‐TAA was restricted to MoMuLV‐transformed cells.
A monoclonal antibody detects heterogeneity in vascular endothelium of tumours and normal tissues
Authors: Wang J. M. , Kumar S. , Pye D. , Krupinski J. , Hunter R. D. , Van Agthoven A. J.
Published: February 2007

A new murine monoclonal antibody (MAb), E‐9, has been raised using tissue‐cultured human umbilical vein endothelial cells. The antigen recognized by this MAb is a peptide of 170 kDa under non‐reducing conditions and 96 kDa under reducing conditions. MAb E‐9 showed marked heterogeneity in its distribution in various tissues. The antigen recognized by it was present in vascular endothelial cells of all tumours, foetal organs and in regenerating and inflamed tissues. It stained a few normal tissues. However, with the exception of tonsils, staining tended to be weak and limited to a few blood vessels, as revealed by double staining using pan‐endothelial antibody (CD31) and antibody to von Willebrand factor, another marker of vascular endothelium. Surprisingly, blood vessels within the placental villi were completely negative. The function of the antigen recognized by MAb E‐9 is not known, but its evaluation and use should increase our understanding of angiogenesis.
Tumour spectrum in hereditary non‐polyposis colorectal cancer (HNPCC) and in families with “suspected hnpcc”. A population‐based study in northern Italy
Authors: Benatti Picro , Sassatelli Romano , Roncucci Luca , De Leon Maurizio Ponz , Pedroni Monica , Fante Rossella , Gregorio Carmela Di , Losi Lorcna , Gelmini Roberta
Published: February 2007

Hereditary non‐polyposis colorectal cancer (HNPCC or Lynch syndrome) is characterized by the early onset of colorectal neoplasms, frequently localized in the right colon, increased occurrence of multiple primaries, vertical transmission and aggregation of tumours in families in accordance to a Mendelian dominant type of inheritance. The syndrome accounts for approximately 5% of all colorectal cancers. The purpose of the present study was to describe the tumour spectrum and the most relevant clinical features of 28 kindreds with HNPCC, classified according to the guidelines of the International Collaborative Study Group, and of 61 “suspected” HNPCC. These families were observed during a 6‐year registration of colorectal neoplasms in a health‐care district of Northern Italy. Colorectal cancer was by far the most frequent malignancy; gastric cancer was the second. Uterine carcinoma was only slightly more frequent than expected. Lung‐ and breast‐tumour rates were lower than expected. Cancer distribution in the large bowel showed that about two fifths of the tumours developed in the right colon. The occurrence of cancer before the age of 50 to 60 was much more frequent in HNPCC. Multiple tumours developed in 25 patients with HNPCC and in 32 with “suspected” HNPCC. Pancolonoscopy remains the procedure of choice for surveillance; other examinations, such as gastroscopy, gynaecological investigations, urography and cholangiography, are suggested only to selected families. One of the main features of the study was the inclusion of 61 “suspected” HNPCC, a heterogeneous group of families which nonetheless deserves careful follow‐up.
Amplification of fgfr4 gene in human breast and gynecological cancers
Authors: Jaakkola Susanna , Salmikangas Paula , Nylund Stella , Lehtovirta Pentti , Nevanlinna Heli , Partanen Juha , Armstrong Elina , Pyrhönen Seppo
Published: February 2007

We have investigated gene amplification of fibroblast growth factor receptor‐4 (FGFR4) gene in 30 primary breast tumor samples and 15 gynecological tumor samples. Ten percent of the breast tumors showed 2‐ to 4‐fold amplification. Amplification was found more frequently in estrogen‐ and progesterone‐receptor‐positive tumors and in tumors with high lymph‐node involvement. Breast tumor samples were also analyzed for the amplification of fgfr3 and erbB2 genes and the chromosome I Iq 13 located genes hst I Iint2lbci1 /sea. erbB2 gene was amplified 2‐ to 13‐fold in 13% of the cases, but no amplification of int2/hst I/be/1/sea amplicon was found. Gynecological tumors were also analyzed for the amplification of fgfr4 and fgfr3 genes and for int2 and hst I oncogenes. Eleven of the 15 gynecological tumors were ovarian neoplasms including 2 benign tumors; the remainder comprised I ovarian metastasis of breast cancer; I endometrial cancer; I uterine leiomyosarcoma and I carcinosarcoma of the fallopian tube. In gynecological tumors, fgfr4 gene was found to be amplified in 2 ovarian tumors. Amplification of hst I was found in I benign ovarian tumor. Thus, the fgfr4 gene may be involved in breast and ovarian tumorigenesis.
Differential chemosensitivity of local and metastatic human gastric cancer after orthotopic transplantation of histologically intact tumor tissue in nude mice
Authors: Furukawa Toshiharu , Kubota Tetsuro , Watanabe Masahiko , Kuo Tsong‐Hong , Kitajima Masaki , Hoffman Robert M.
Published: February 2007

We have established a metastatic model of human gastric cancer using orthotopic transplantation of histologically intact tissue in nude mice, and have used this model to evaluate the effects of immunochemotherapy using OK‐432, 5‐fluorouracil (5‐FU) and mitomycin C (MMC) against SC‐I‐NU, a human stomach cancer line. One‐quarter or one‐half maximum tolerated doses (MTDs) of S‐FU or MMC resulted in a significant reduction of stomach tumor growth, while liver metastases were not reduced, possibly due to suppression of natural killer (NK)‐cell activity by both drugs. On the other hand, when combined with OK‐432, half MTDs of 5‐FU and MMC significantly reduced liver metastases, with synergistic reduction of stomach tumor growth, possibly reflecting a rescue of NK‐cell activity by treatment with OK‐432. This metastatic model of human stomach cancer shows that locally growing and metastatic tumors may have different chemosensitivities, and provides the opportunity to test both with various treatment regimens.
Generation and control of metastasis in experimental tumor systems; inhibition of experimental metastases by a tilorone analogue
Authors: Algarra Ignacio , Pérez Matías , Garrido Federico , Gaforio Jose Juan , Höglund Petter , Ljunggren Hans‐Gustaf
Published: February 2007

The role of the chemical compound RMI 10,874DA (3,6‐bis[2‐(dimethylamino)‐ethoxyl]‐9H‐xanthene‐9‐onedihydrochloride) in the abrogation of the metastatic spread of tumor cells was studied. Pre‐treatment of BALB/c mice with the RMI 10,874DA compound (referred to below as tilorone analogue) completely eliminated lung colonization of an H‐2‐negative (GR9.B9) MCA‐induced fibrosarcoma clone in an experimental metastasis assay. Other murine tumors, including H‐2‐positive and H‐2‐negative chemically induced fibrosarcoma clones and B16 melanoma, were also sensitive to the treatment; orally administered tilorone analogue given one day before the i.v. injection of tumor cells markedly inhibited lung colonization. The effect was not due to direct toxicity of tilorone analogue on tumor cells, but instead it was dependent on NK cells; this was suggested by the finding that anti‐asialo GM1 treatment of mice abrogated the effect of tilorone analogue. Kinetic studies of splenic NK activity in tilorone‐treated mice showed a rapid boosting of NK‐cell activity, the greatest stimulation occurring the day before removal of splenocytes for 51Cr‐release assay against YAC‐1 target cells. These kinetics correlated with the inhibition of in vivo lung colonization after tilorone analogue treatment. Inhibition of experimental tumor metastasis was dose‐dependent and was observed when animals were treated the day before or the day after tumor‐cell injection. Furthermore, repeated treatment of mice with this tilorone analogue significantly reduced lung colonization.
Tumor‐suppressor p53 gene in hepatitis C and B virus‐associated human hepatocellular carcinoma
Authors: Carol Shieh Y. S. , Nguyen Chau , Vocal Maria V. , Chu Hoi‐Weng
Published: February 2007

Abnormalities of the tumor‐suppressor pS3 gene have been discovered in human hepatocellular carcinoma (HCC). It is unclear, however, whether HCC related to chronic viral hepatitis is associated with p53 gene alterations. In this study, we have examined pS3 abnormalities in HCC associated with hepatitis C and B virus (HCV and HBV) infections. Tissues from 18 HCC patients from several hospitals throughout the United States were collected (9 were HCV‐infected, 5 were HBV‐infected, I was HCV/HBV‐infected, and 3 were non‐virus‐associated). Immunostaining with monoclonal pAb1801 revealed expression of p53 protein in tumor‐cell nuclei in one HCV‐associated HCC, and in no case of HBV‐associated HCC, while the nuclei of adjacent hepatocytes were negative. Using Hae Ill‐digestion of chromosomal DNA, mutations at codon 249 were not found in any of 18 HCC tissues studied. Direct sequencing demonstrated a mutated codon 244 and a wild‐type codon 249 in the conserved regions (exon 5–8) of p53 gene from the tumor tissue with nuclear p53 expression. By reverse‐transcription‐polymer‐ase chain reaction (RT‐PCR), the expression of p53 mRNA was demonstrated in tumor cells from 10 out of 16 HCC tissues. In conclusion, the specific mutation at codon 249 with G to T transversion was not observed in the HCCs associated with HCV or HBV infections. In HBV or non‐virus‐associated HCCs studied, no other p53 gene abnormalities were found. A point mutation at codon 244 with G to A transition of p53 gene was detected in only one of 10 HCV‐associated HCCs, which suggests that p53 mutations may not play a significant role in HCV‐ or HBV‐associated hepatocarcinogenesis.
Novel protein kinases expressed in human breast cancer
Authors: Cance William G. , Craven Rolf J. , Weiner Timothy M. , Liu Edison T.
Published: February 2007

The family of protein kinases includes many oncogenes and growth‐factor receptors, as well as genes that are involved in cell‐cycle regulation. We have identified protein kinases expressed in a human breast‐cancer cell line, 600PEI, and a primary human breast carcinoma, using PCR cloning techniques based on consensus sequences in the kinase domain. Twenty‐five different protein kinases were isolated, including 3 novel putative tyrosine kinases (designated TKI, TK2, and TK5), and 2 novel putative cell‐cycle‐associated serine/threonine kinases (designated STKI and STK2). TKI is a new member of the src family of kinases that is expressed predominantly in epithelial cells. TK2 is homologous to the receptor kinase, HEK, and TK5 appears to be another member of the J AK family of kinases. The novel serine/threonine kinases, designated STKI and STK2, were homologous to the human cdc2 and the Aspergillus nimA genes. We subsequently analyzed the levels of expression of all of these protein kinases in a panel of human breast carcinomas, using PCR‐based methods. This analysis revealed different expression profiles in different primary breast carcinomas and, therefore, may determine new molecular sub‐sets of human.
Malignant mesothelioma after environmental exposure to blue asbestos
Authors: Hansen Janice , De Klerk Nicholas H. , Eccles Jan L. , William Musk A. , Hobbs Michael S. T.
Published: February 2007

To determine the magnitude of the population at risk from non‐occupational exposure to crocidolite at Wittenoom, West ern Australia (WA), a cohort of 4,890 residents who never worked for the mining company Australian Blue Asbestos (ABA) has been assembled from all 18,553 available records: the local school register, hospital attendances, the WA electoral roll, birth certificates, workers who answered a mailed question naire in 1979, participants in a cancer‐prevention programme using vitamin‐A dietary supplements, and other sources. The majority of subjects were relatives and friends of ABA employ ees, and nearly half the cohort were either born at Wittenoom or first went there as children under 10 years of age. As most residents were at Wittenoom when the mine and mill were in operation during the period 1943 to 1966, 82% were first exposed to crocidolite 20 or more years ago. The proportion of other workers (i.e., not employed by ABA) and their families increased once the mining operations ceased. To date, 24 cases of mesothelioma have been reported in this cohort: 9 males and 15 females. Time from first exposure to diagnosis ranged from 23 to 44 years and residence in Wittenoom ranged from 6 weeks to 11 years.
X‐ray‐induced chromatid damage in relation to dna repair and cancer incidence in family members
Authors: Knight Robert D. , Parshad Ram , Price Floyd M. , Tarone Robert E. , Sanford Katherine K.
Published: February 2007

The cytogenetic response to G2‐phase X‐irradiation was examined in phytohemagglutinin‐stimulated peripheral‐blood lymphocytes from 69 individuals, a few of whom were cancer patients. The cancer patients had not received radiation or chemotherapy. The responses of cells arrested by Colcemid 30 to 90 min after X‐irradiation (58R) could be divided into 2 distinct categories: 51 individuals had aberration frequencies typical of normal individuals in previous studies, while 18 others had a 2‐ to 3‐fold higher frequency of chromatid breaks and gaps. Because chromatid breaks and gaps result from unre paired DNA strand breaks, the first category may represent an efficient DNA repair phenotype, while the second may repre sent a deficient repair phenotype. The individuals with the deficient G2 response reported having first‐ and second‐degree relatives with a 3.6‐ and 2.2‐fold higher mean frequency of cancer, respectively. The present results, together with those from earlier studies of families with a genetic disorder predispos ing to cancer, suggest that this deficient cytogenetic response to G2 phase X‐irradiation is associated with a high risk of cancer.
Prediction of response to endocrine therapy in breast cancer using immunocytochemical assays for pS2, oestrogen receptor and progesterone receptor
Authors: Luqmani Y. A. , Ricketts D. , Ryall G. , Turnbull L. , Law M. , Coombes R. C.
Published: February 2007

Histological sections obtained from 70 patients with breast cancer, all of whom had received endocrine therapy for meta‐static or locally advanced disease, were assessed for specific immunocytochemical staining of oestrogen receptor and the oestrogen‐induced protein pS2. There was also sufficient material from 25 patients for an assessment of progesterone receptor by immunocytochemistry. We found that, when using a „cut‐off”︁ point of 50% for ER and PR, and of 25% for pS2, ER was positive in 22/29 responders and in 12/41 non‐responders, and thus was significantly associated with response to endocrine therapy. Similarly, in those subjects in whom PR was measured, PR was positive in 5/14 responders and negative in all II non‐responders, again being significantly correlated with response. However, pS2 did not relate to response, being only positive in 10/29 responders and negative in 26/41 non‐responders. The different response categories varied in their „percentage of positivity”︁ as determined by the 2 tests. Thus, for ER and pS2 we observed: complete response—71 % for ER compared with 14% for pS2; partial response—77% compared with 41%; stable disease—36% compared with 64%; and progressive disease—27% compared with 27%. We conclude that at the present time ER appears to be the most reliable indicator for predicting response to endocrine therapy in patients with breast cancer.
Lysis of human pancreatic adenocarcinoma cells by autologous hla‐class I‐restricted cytolytic T‐lymphocyte (CTL) clones
Authors: Wölfel Thomas , Herr Wolfgang , Couue Pierre , Schmitt Ute , Zum Büschenfelde Karl‐Hermann Meyer , Knuth Alexander
Published: February 2007

From the primary site of a pancreatic adenocarcinoma (patient BE) a permanent cell line (MZ‐PC‐2) was established in tissue culture. In the course of mixed lymphocyte‐tumor‐cell cultures (MLTC) with autologous blood‐derived lymphocytes, we isolated CTL clones that lysed autologous tumor cells but not autologous EBV‐transformed B cells (EBV‐B) and not K562. Pre‐treatment of MZ‐PC‐2 cells with IFN‐γ was required to obtain significant lysis in 4‐hr cytotoxicity assays. IFN‐γ was superior to IFN‐α in that respect. Among MLTC responder lymphocytes, tumor‐reactive CTL proliferated more strongly in response to MZ‐PC‐2 cells treated with IFN‐γ than to untreated tumor cells. Three CTL clones derived from MLTC were chosen for further analysis. They were CD3+, CD8+, TCR‐α/β+ and behaved identically in all functional aspects tested. They all expressed the same TCR‐β chain, indicating that they descended from a common precursor lymphocyte and were directed against the same antigen. According to antibody‐inhibition experiments, BE‐CTL recognized their targets via an HLA‐B molecule carrying the Bw6 supertypic determinant. Irrespective of pre‐incubation with IFN‐γ, low levels of tumor‐cell lysis, or none, were seen when MZ‐PC‐2 cells were kept in medium supplemented with autologous serum or serum pooled from healthy volunteers instead of FCS. Lysability was restored when TNF‐α was added to human serum. Serum‐free medium was found to enhance the susceptibility of MZ‐PC‐2 cells to lysis by autologous CTL.
Role of plasminogen activators, metalloproteinases and the tissue inhibitor of metalloproteinase‐1 in the metastatic process of human salivary‐gland adenocarcinoma cells
Authors: Azuma Masayuki , Tamatani Tetsuya , Fukui Kazuhiro , Yoshida Hideo , Kamogashira Takashi , Ogino Koichi , Suzuki Toshimitsu , Sato Mitsunobu
Published: February 2007

An in vitro system has been established in which conversion from non‐metastasizing to metastasizing adenocarcinoma cells can be induced, and subsequently subjected to analysis of the expression of proteases and tissue inhibitor of metalloprotein‐ases‐l (TIMP‐I). A human salivary‐gland adenocarcinoma cell clone HSGc, with no metastatic ability, was exposed to N‐meth‐yl‐N‐nitrosourea (MNU). Following exposure to MNU, cells with altered morphology were cloned. Upon s.c. inoculation into nude mice, MNU‐treated HSGc clones formed metastatic foci in various organs, and then 5 metastasizing clones were isolated. Evaluation of expression of tissue‐type plasminogen activator (tPA), urokinase‐type plasminogen activator (uPA), metalloproteinases and TIMP‐I was performed by means of enzyme immunoassay, zymogram, or immunoblot. MNU‐treated HSGc and metastasizing clones were found to secrete high levels of tPA, while HSGc produced undetectable levels of this enzyme. Expression of uPA was not observed in any of the cell clones. When the secretion of gelatinolytic enzymes was examined, metastasizing clones produced higher levels of 57–and 32–kDa, but not of 92– or 72–kDa gelatinases, as compared to HSGc cells. Although TIMP‐I was detected in all cell clones, metastasizing clones secreted less TIMP‐I than HSGc cells; in addition, one metastasizing clone produced TIMP‐I with a molecular weight distinct from that of 28–kDa TIMP‐I. Our results suggest that the acquisition of metastatic ability by human salivary‐gland tumor cells is closely associated with increased secretion of several metalloproteinases as well as decreased or altered TIMP‐1 expression.
Identification of colon‐tumor‐associated antigens by T‐cell lines derived from tumor‐infiltrating lymphocytes and peripheral‐blood lymphocytes from patients immunized with an autologous tumor‐cell/bacillus calmette‐Guérin vaccine
Authors: Ransom Janet H. , Pelle Barbara A. , Hubers Henk , Keynton Linette M. , Hanna Michael G. , Pomato Nicholas
Published: February 2007

Tumor immunity developing as a response to an autologous colon‐tumor/bacillus Calmette‐Guerin (BCG) vaccine appears to be associated with induction of CD4 helper T cells, implied by the observation that vaccine efficacy is associated with major histocompatibility complex class‐ll molecule expression on the vaccine tumor cells. Therefore, in an attempt to identify colon‐tumor‐associated antigens responsible for conferring immunity, we examined and compared the proliferative responses of peripheral‐blood lymphocytes (PBL) from patients immunized with the autologous tumor/BCG vaccine to T‐cell lines cloned expanded from colon‐tumor‐infiltrating lymphocytes to 5 antigens isolated on the basis of their reactivity by colon‐tumor‐reactive human monoclonal antibodies. Enzymatically dissociated colon tumors provided a source for establishment of cloned T‐cell lines, tumor cell lines propagated in vitro or in vivo as nude‐mouse xenografts and EBV‐transformed B‐cell lines used as antigen‐presenting cells. Of 104 different T‐cell lines tested, only 3 proliferated in response to CTAA 28A32‐46K, and I to the CTAA28A32‐32K antigen. In contrast, PBL from 64% of patients immunized with the autologous colon‐tumor/BCG vaccine responded to the CTAA 28A32‐32K antigen. This antigen is related to a family of calcium‐ and phospholipid‐binding placental proteins termed annexins. Since proliferative responses developed to this antigen after vaccination in 64% of individuals, this antigen may be an important common colon‐tumor‐associated rejection antigen.
Influence of interleukin‐2 administration on the expression of T‐cell receptor V gene segments in patients with renal‐cell carcinoma
Authors: Farace Françoise , Angevin Eric , Escudier Bernard , Caignard Anne , Dietrich Pierre‐Yves , Genevee Catherine , Hercend Thierry , Triebel Frédéric
Published: February 2007

Tumor regression induced by IL‐2 in a fraction of patients with metastatic renal‐cell carcinoma (MRCC) could not be predicted by immunological monitoring. In addition, the general mechanisms leading to tumor regression or even the distinct cell subsets (e.g., T vs. NK cells) involved are poorly identified. To evaluate the influence of IL‐2 administration on circulating T‐cell subpopulations, TCR Vα and Vβ gene segment usage was analysed by PCR in 7 MRCC patients using a panel of V gene segment subfamily‐specific oligonucleotide primers (Vα‐w29/ VβI‐w24). Three samples were examined in each patient: (i) peripheral blood cells (PBMCs) before therapy (day I); (ii) PBMC 2 days after the interruption of IL‐2, at day 36 (i.e., at the lymphocytic rebound), (iii) the CD25‐enriched cell fraction at day 36. Virtually all Vα and Vβ subfamily specificities were found in pre‐ as well as in post‐treatment PBMCs and CD25+ cell fractions. These results support the view that circulating T‐cell subpopulations are highly polyclonal after IL‐2 therapy without any major alteration in the TCR Vα and Vβ repertoire. In addition, the results of quantificative densitometric analysis of Vα and Vβ amplified products suggest that a unique Vβ‐expressing T‐cell subpopulation may be expanded in the CD25+ cell fraction after IL‐2 therapy. Further characterization of these T cells may contribute to a better understanding of in vivo effects of IL‐2 on renal‐cell carcinoma metastases.
Deletions of the shorlt arm of chromosome 9, including the interferon ‐ α / β genes, in acute lympocytic leukemia. Stuides on loss of heterozygosity, parental origin of deleted genes and prognosis
Authors: Heyman Mats , Grai Dan , Bröndum‐Nielsen Karen , Lir Yie , Söderhäll Stefen , Einhorn Stefen
Published: February 2007

A restriction fragment length polymorphism (RFLP) analysis of the α‐ and β‐interferon (IFN) genes was performed in malignant cells from 52 patients with acute lymphocytic leukemia (ALL). Normal cell DNA was available for comparison in 23 of the patients. Ten patients were found to have gross alterations of their α‐ and β‐IFN genes. Leukemic cells from 2 ALL patients showed a complete loss of α‐ and β‐IFN genes. Seven patients had a hemizygous loss of one of the α‐ and β‐IFN alleles, as shown by RFLP analysis and/or loss of signal intensity. In one other patient the malignant clone was reduced to homozygosity with regard to the α‐ and β‐IFN genes, without loss of signal intensity. In patients without hemizygous deletions, the overall incidence of complete homozygosity for the α‐ and β polymorphisms was higher than expected. Analysis of the data indicates that the total frequency of ALL clones with gross alterations of the IFN‐loci is around 30%. A 9p24 probe detected hemizygous deletions in 2 cases of IFN gene deletions. In the other tested cases the deletions were interstitial. No deletions of 9p24 were detected in patients without allelic losses of IFN genes. In 5 cases of allelic IFN gene deletions, DNA from parents was available for comparison. In 4 cases the deleted allele was derived from the mother, whereas in the fifth it originated from the father. Pediatric ALL patients with IFN‐gene deletions or homozygosity for all polymorphisms in the IFN‐loci had a significantly worse prognosis than heterozygotes. We conclude that deletion of α‐ and β‐IFN genes is a relatively common event in ALL and that RFLP analysis of the IFN genes may provide additional prognostic information in childhood ALL. Whether or not the IFNs act as tumor‐suppressor genes in this disease is not yet known. (c) 1993 Wiley‐Liss, Inc.
Acidic fgf and egf are involved in the autocrine growth stimulation of a human nasopharyngeal carcinoma cell line and sub‐line cells
Authors: Chao Hsiou‐Hsien , Yang Vivian C. , Chen Jan‐Kan
Published: February 2007

The growth of a recently established human nasopharyngeal carcinoma cell line, CG‐1, and 5 randomly selected, single‐cell‐derived sub‐lines in serum‐free medium and in fetal‐bovine‐serum (FBS)‐containing medium was investigated. In basal medium supplemented with insulin, transferrin, fibronectin and high‐density lipoprotein, cell growth was moderately stimulated by aFGF and EGF in a dose‐dependent manner. In contrast, in medium containing as little as 0.5% FBS, most of the stimulatory effect of the aforementioned growth factors observed was masked. Western blotting analysis of the cell lysates and conditioned media showed that CG‐1 and sub‐line cells were all capable of synthesizing and releasing aFGF‐ and EGF‐immunoreactive proteins. The amounts of these 2 growth factors synthesized and released appeared to vary among the parental cell line and sub‐line cells. Moreover, the rate of basal proliferation of these cells appeared to be positively correlated with the amounts of aFGF‐ and EGF‐immunoreactive proteins produced. Addition of the neutralizing antibodies to aFGF and EGF exerted a dose‐dependent suppression on cell growth in medium containing 0.5% FBS. The results suggest a role of aFGF and EGF in autocrine growth stimulation of CG‐1 and sub‐line cells, and may explain the moderate response of these cells to exogenously added aFGF and EGF.
Differential metabolism and trafficking of sphingolipids in differentiated versus undifferentiated HT29 cells
Authors: Babia Teresa , Kok Jan Willem , Hulstaert Caesar , De Weerd Henk , Hoekstra Dick
Published: February 2007

Trafficking and metabolism of sphingolipids were examined in undifferentiated (G+) and differentiated (G+ reversed) HT29 human colon adenocarcinoma cell lines. Metabolic experiments employing a fluorescently labeled sphingolipid precursor, 6‐[N‐(7‐nitro‐2,1,3‐benzoxadiazol‐4‐yl)amino]hexanoylceramide (C6‐NBD‐ceramide) revealed that both qualitative and quantitative differences exist in sphingolipid synthesis between the 2 cell lines. One of the C6‐NBD‐sphingolipids synthesized in G+ cells is not found in the G+ reversed cells. Furthermore, the ratio of the 2 main products, C6‐NBD‐glucosyiceramide and C6‐NBD‐sphingomyelin, differs: in G+ cells glucosylceramide is by far the main product, whereas G+ reversed cells synthesize C6‐NBD‐sphingomyelin in slight excess. Once established, these ratios of sphingolipids are quickly restored metabolically when distortion of the ratio is caused by experimental manipulation. This indicates that they represent a true metabolic equilibrium situation of the 2 sphingolipids in these cells, while the distinct ratios are mainly determined by the NBD‐lipid pool in the plasma membrane. Preferential synthesis and transfer of glucosylceramide from its site of synthesis to the cell surface do not occur when the plasma membrane pool of glucosylceramide is selectively removed. This suggests that instantaneous replenishment via specific signalling is probably not involved as a mechanism in re‐establishing perturbed lipid pools. In conjunction with observations on distinct lipid trafficking pathways of glucosylceramide in G+ and G+ reversed cells, the present metabolic studies emphasize a relation between the expression of this glycolipid and the state of differentiation of HT29 cells.
A novel monoclonal antibody specifically reactive with human T‐lymphotropic virus type‐II (HTLV‐II) envelope protein
Authors: Ohara Nobuya , Hayashi Kazuhiko , Takahashi Kiyoshi , Miyamoto Kanji , Akagi Tadaatsu
Published: February 2007

A novel monoclonal antibody (MAb), N5.4.4, specifically reactive with human T‐lymphotropic virus type‐ll (HTLV‐II) envelope glycoprotein (gp) has been developed through immunization with a synthetic peptide corresponding to the amino acid sequence 171–196 of the HTLV‐II envelope protein (ll‐env 171–196). This MAb, which belonged to the IgG1 kappa subclass, reacted with the cytosmears of HTLV‐ll‐infected cell lines (Si‐IIA, CR‐MA‐I and AS‐IIA), but not with those of HTLV‐1‐infected cell lines (MT‐1 and MT‐2) or other HTLV‐uninfected cell lines. On Western blot analysis, this MAb reacted with gp46 of HTLV‐II lysates but not with HTLV‐1 lysates. Moreover, flow‐cytometric analysis revealed that this MAb recognized the native surface of only the HTLV‐ll‐producing cells. Through application to immunohistochemical or serological method, this MAb may be of value in elucidating the pathogenesis of HTLV‐II infection in comparison with HTLV‐1.
Induction of mouse tenascin expression by a human sarcomatoid wilms' tumor cell line growing in nude mice
Authors: Talts Jan Fredrik , Aufderheide Enno , Sorokin Lydia , Ocklind Göran , Mattsson Ragnar , Ekblom Peter
Published: February 2007

A new cell line from a sporadic Wilms' tumor was established and extensively characterized. In nude mice, the tumor cells rapidly formed tumors which, in histological characteristics and extracellular‐matrix (ECM) composition resembled sarcomatoid Wilms' tumor. The tumor cells produced B chains of laminin, but no A chain, and laminin was deposited into the ECM in a punctate pattern typical of sarcomatoid tumors. Strong expression of tenascin was detected within the stromal ECM of the tumors. Species‐specific antibodies reacting either with human or with mouse tenascin showed that tenascin was exclusively derived from mouse host cells. The human Wilms' cell line thus induced a strong stromal response with increased deposition of tenascin. The cell line may be useful for studying the behavior of sarcomatoid Wilms' tumor cells and for identifying factors that stimulate synthesis of tenascin.
Cancer incidence in the native peoples of far eastern siberia
Authors: Zaridze David G. , Marochko Andrej , Basieva Tamara , Duffy Stephen W.
Published: February 2007

Cancer incidence rates were examined in the native peoples of the far north‐east of Siberia for the years 1977–1988. Particularly high rates of cancers of the stomach, lung, oesophagus and cervix were observed. For stomach cancer, the male and female age‐standardized (to the world population) rates were 103.9 per 100,000 and 50.0 per 100,000 respectively. The corresponding lung cancer rates were 109.4 and 45.7, and for oesophageal cancer 83.9 and 35.0. The age‐standardized cervical cancer rate was 38.5 per 100,000. Rates of these cancers were considerably higher than in native Alaskan peoples, although the latter had higher rates of breast and colorectal cancers. The rates were also much higher than those of the migrant peoples from Russia and elsewhere who have settled in the far north‐east over the past 3 centuries, particularly at younger ages. Male rates of stomach and lung cancer were highest in the paleo‐Asiatic peoples of the north, whereas male oesophageal rates were highest in the Taiga people. In females, rates of stomach and oesophageal cancers were highest in the paleo‐Asiatic peoples, and rates of lung cancer were highest in the Taiga nationalities. Cervical cancer rates were highest in the Amuro‐Sakhalin nationalities of the south. Further research is needed at individual levels to explain the very high risks and the differences among the ethnic groups.
Parity and cancers of the gall bladder and the extrahepatic bile ducts
Authors: Lambe Mats , Trichopoulos Dimitrios , Hsieh Chung‐Cheng , Ekbom Anders , Adami Hans‐Olov , Pavia Maria
Published: February 2007

The relation of parity and age at first birth to cancers of the gall bladder and extrahepatic bile ducts in women was studied using a database generated by linking 2 Swedish national registries; the Fertility Registry and the Cancer Registry. Among women born between 1925 and 1960, 257 cases of gall‐bladder cancer recorded in the Cancer Registry between 1958 and 1984 were compared with 1,285 controls, age‐matched to cases in a 5:1 ratio. In addition, 60 cases of extrahepatic‐bile‐duct cancer were matched with 300 controls. There was a positive association between number of live births and risk of gall‐bladder cancer (p 0.06), but simultaneous consideration of parity and age at first birth revealed a more complex picture. Parity increases the risk for cancer of the gall bladder when the first birth occurs before the age of about 25 years, whereas parity associated with first birth after the age of about 30 years is associated with reduced risk for the disease. Thus, among parous women there is a highly significant inverse association of age at first birth with risk for gall‐bladder cancer after adjustment for number of live births. Variable levels of pregnancy estrogens according to maternal age and variable effects of parity on non‐pregnancy estrogens by age, may explain the observed pattern. The results on extrahepatic‐bile‐duct cancer, parity and age at first birth did not indicate the existence of an association in either direction.
Modulating the metastatic potential of murine raw117 large‐cell lymphoma cells by selection for resistance to interferon‐γ
Authors: Labiche Ronald A. , Nicolson Garth L.
Published: February 2007

Highly metastatic, in vivo‐selected cells of RAW117‐H10 large‐cell lymphoma have been shown to be more resistant than poorly metastatic parental RAW117‐P cells to the cytolytic and cytostatic activities of activated macrophages in co‐culture experiments. Activated macrophages are known to produce soluble, cytostatic respiration‐inhibiting factors, and such activities can be duplicated by interferon‐gamma (IFN‐γ) or by combinations of IFN‐γ and Escherichia coli lipopolysaccharide (LPS). Highly metastatic RAW117‐H10 cells are more resistant to the cytostatic effects of IFN‐γ and LPS than poorly metastatic RAW117‐P cells, and short‐term (up to 72 hr) treatment with IFN‐γ and LPS does not change the metastatic potentials of RAW117 cells. We have studied the effects of sequential selection of RAW117‐P cells for increased resistance to IFN‐γ and LPS. After 7 to 13 sequential selections, the resulting variant lines were completely refractory to the growth‐inhibitory effects of IFN‐γ and LPS and cross‐tolerant to inacro‐phage‐conditioned medium. The selected variants also completely lost their experimental metastatic potentials and their tumorigenicities after s.c. or i.m. injection. Cytofluorographic analysis indicated reduced cell‐surface expression of H‐2Kd antigen and fibronectin receptor on the variant cells but no change in surface μ heavy‐chain immunoglobulin. The IFN‐γ‐selected lines were less adhesive to liver microvascular endothe‐lial cells than the unselected RAW117 cell lines, but were equally sensitive to NK cytolysis by spleen cells. Our results suggest that exposure to certain cytokines can affect the growth and metastatic potential of RAW117 cells and result in the selection of resistant variants with altered biologic properties.
Role and regulation of expression of 92‐kDa type‐IV collagenase (MMP‐9) in 2 invasive squamous‐cell‐carcinoma cell lines of the oral cavity
Authors: Juarez Jose , Clayman Gary , Nakajima Motowo , Tanabe Kenneth K. , Saya Hideyuki , Nicolson Garth L. , Boyd Douglas
Published: February 2007

The present study was undertaken to determine the role of the metalloproteinase MMP‐9 in the invasive phenotype of squamous‐cell carcinoma of the oral cavity and the regulation of its expression. Zymographic analysis of conditioned medium from 2 highly invasive squamous‐cell‐carcinoma cell lines indicated large amounts of an enzyme which was indistinguishable, in size (92 kDa) from the MMP‐9 pro‐enzyme. Conversion of the 92‐kDa gelatinase into a lower‐molecular‐weight species (84 kDa), identical in size to the activated gelatinase, was evident when both cell lines, which are avid secretors of urokinase, were cultured in the presence of plasminogen. Penetration of an extracellular‐matrix‐coated filter was dramatically reduced in the presence of the collagenase inhibitor, tissue inhibitor of metalloproteinase‐2, suggesting a critical role for MMP‐9 in the invasive process. Immunohistochemical studies demonstrating the presence of MMP‐9 in tumor cells of resected squamous‐cell cancers suggested that secretion of this collagenase by cells in vitro was reflective of the in vivo setting. Since several phorbol‐ester response elements are present in the MMP‐9 promoter, we determined the role of protein‐kinase‐C pathways in the regulation of MMP‐9 expression in cultured SCC. Treatment of cells with PMA resulted in a more‐than‐20‐fold increase in the level of protein and mRNA. Conversely, culturing of cells in the presence of the protein‐kinase‐C inhibitor, calphostin‐C, led to a dose‐dependent decrease in the amount of MMP‐9 mRNA and protein, suggesting that the constitutive expression of this collagenase reflects activation of this signal transduction pathway. In summary, our data suggest that, for a sub‐population of squamous‐cell carcinomas, secreted MMP‐9 is an important determinant of the invasive phenotype, and that the expression of this metalloproteinase is regulated by protein‐kinase‐C pathways. © 1993 Wiley‐Liss, Inc.
The plasminogen‐activation system in ovarian tumors
Authors: Pujade‐Lauraine E. , Lu H. , Mirshahi S. , Soria J. , Soria C. , Bernadou A. , Kruithof E. K. O. , Lijnen H. R. , Burtin P.
Published: February 2007

We studied the plasminogen activation system in tumor tissue by measuring the antigen level of the 2 plasminogen activators, tissue‐type (t‐PA) and urokinase‐type (U‐PA) and their inhibitors, plasminogen‐activator inhibitors type‐1 (PAI‐1) and type‐2 (PAI‐2) in the tissue extracts of 43 human benign and malignant ovarian tumors. U‐PA levels were significantly higher in malignant than in benign tumors. In addition, U‐PA antigen levels were higher in the metastatic tissue of advanced disease (FIGO stage III) than in the primary localized tumor (FIGO stage I/II). Also PAI‐1 concentrations tended to be higher in malignant than in benign tumors, but this difference was not statistically significant. In contrast, t‐PA levels were lower in metastatic than in non‐metastatic tumors, whereas PAI‐2 levels were unrelated to the stage of ovarian malignancy. These results were integrated in a plasminogen‐activation‐dependent malignancy index (U‐PA × PAI‐1/t‐PA). This index distinguished the different groups of benign ovarian tumors, localized and metastatic ovarian carcinomas better than U‐PA levels. It could be useful as a prognostic indicator in ovarian cancer © 1993 Wiley‐Liss, Inc.
Aberrant expression of c‐met mRNA in human gastric carcinomas
Authors: Kuniyasu Hiroki , Yasui Wataru , Yokozaki Hiroshi , Kitadai Yasuhiko , Tahara Eiichi
Published: February 2007

We examined mRNA expression for c‐met encoding hepatocyte growth factor receptor in 8 gastric carcinoma cell lines and 31 surgically resected gastric carcinoma tissues by Northern‐blot analysis. Two forms of the transcript, sized 7.0 kb and 6.0 kb, were found in gastric carcinomas. Transcripts of both sizes were detected at various levels in all the gastric carcinoma cell lines except the MKN‐74 line, which expressed only the 7.0‐kb form. Of the 31 gastric carcinomas, 15 (48%) over‐expressed the 7.0‐kb transcript when compared with non‐neoplastic mucosa. The 6.0‐kb transcript of the c‐met gene was expressed at considerable levels in 52% of the gastric carcinoma tissues, whereas it was only slightly expressed in non‐neoplastic mucosa of a small number of cases. An important finding was that expression of the 6.0‐kb transcript was closely correlated with tumor staging, lymph‐node metastasis and depth of tumor invasion. These results, overall, suggest that the 6.0‐kb transcript of the c‐met gene might participate in the development and progression of gastric carcinomas. © 1993 Wiley‐Liss, Inc.
Interleukin‐10 production by human carcinoma cell lines and its relationship to interleukin‐6 expression
Authors: Gastl Guenther A. , Abrams John S. , Nanus David M. , Oosterkamp Rianne , Silver John , Liu Frank , Chen Michael , Albino Anthony P. , Bander Neil H.
Published: February 2007

Recent data indicate a major role for IL‐10 in suppressing immune and inflammatory reactions. To date, expression of human IL‐10 has been attributed primarily to helper T lymphocytes, activated monocytes, and neoplastic B cells, and was often found to be associated with IL‐6 expression. In this study we sought to determine whether non‐hematopoietic human tumor cell lines produce IL‐10 and, if so, what is the relationship between IL‐10 and IL‐6. Using ELISA, we determined IL‐10 and IL‐6 levels in culture supernatants of 48 cell lines established from carcinomas of the kidney, colon, breast and pancreas, malignant melanomas and neuroblastomas. IL‐6 protein was secreted by 28 of the tumor ceil lines; IL‐10 was measurable in 15 cell lines. IL‐6 secretion was maximal and most frequent in renal‐cancer cell lines, while IL‐10 production was found to be highest and most common among cell lines derived from colon carcinomas. IL‐10 in conditioned medium of one of the colon carcinoma cell lines (CCL222) was bio‐active, as demonstrated in the mouse MC/9 mast‐cell‐line assay and in human mixed‐lymphocyte reactions. In both assays, IL‐10 bio‐activity was neutralized by an anti‐IL‐10 monoclonal antibody. Expression of IL‐6 and IL‐10 was confirmed by RNA analysis using message amplification by PCR and sequencing of amplified cDNA. LPS, IL‐1 alpha, and TNF‐alpha strongly enhanced the release of IL‐6 by RCC cells, but only marginally affected IL‐10 production in colon‐carcinoma cells. IL‐10 secretion by colon‐carcinoma cells was moderately stimulated by IFN‐gamma and IL‐4. Dexamethasone suppressed the release of IL‐6, but had no inhibitory effect on IL‐10 secretion. Our results demonstrate that tumor cell lines established from certain types of human carcinomas are capable of expressing and releasing IL‐6 and/or IL‐10, suggesting a role of these cytokines in solid‐tumor development and anti‐tumor immunity. © 1993 Wiley‐Liss, Inc.
Differential over‐expression of mdr1 genes in multidrug‐resistant rat glioblastoma cell lines selected with doxorubicin or vincristine
Authors: Schott Brigitte , Bennis Sanae , Pourquier Philippe , Ries Colette , Londos‐Gagliardi Danielle , Robert Jacques
Published: February 2007

We have compared the pharmacological and molecular characteristics of 2 cell lines derived from the C6 rat glioblastoma, and selected for resistance either to doxorubicin (C6 0.5 line) or to vincristine (C6 IV line). Each line displays a preferential 400‐fold resistance towards the drug used for selection, the C6 IV line being especially weakly resistant to doxorubicin (13‐fold). Verapamil completely restored doxorubicin sensitivity in the C6 IV line as well as vincristine resistance in the C6 0.5 line, but could not completely reverse doxorubicin resistance in the C6 0.5 line or vincristine resistance in the C6 IV line. This suggests that specific mechanisms of resistance against each drug were added to a common P‐glycoprotein‐mediated multi‐drug‐resistance mechanism. Doxorubicin efflux was total within 2 hr in the C6 IV line, whereas it remained 8 to 10% of drug in the C6 0.5 line 4 hr after drug removal, despite a more rapid efflux of the drug in the first 30 min. This 2‐compartment behavior could be related to a special sub‐cellular distribution of doxorubicin in C6 0.5 cells. Northern and Western blot analysis of the mdr I gene and of the P‐glycoprotein expressed by the 2 resistant cell lines made it possible to quantify their degree of over‐expression; when compared with the C6 wild strain, the C6 0.5 line over‐expressed both the mdr1 gene and the P‐glycoprotein to a slightly higher level than the C6 IV line. Northern and Western blot analysis also suggested that C6 0.5 cell preferentially over‐expressed the mdr I a gene, whereas the C6 IV cells preferentially over‐expressed the mdr Ib gene. This differential over‐expression was confirmed after polymerase‐chain‐reaction amplification of the cDNA sequences transcribed from total RNA extracted from the 2 lines. It can be concluded therefore that the mdr Ia gene product is more efficient than the mdr Ib gene product in extruding anti‐cancer drugs from the cells; and that the mdr Ib gene product might preferentially extrude vincristine rather than doxorubicin. © 1993 Wiley‐Liss, Inc.