Establishment and characterization of a carcinoembryonic antigen (CEA)-producing cell line from a human carcinoma of the exocrine pancreas

Okabe, Tetsuro; Yamaguchi, Naohito; Ohsawa, Nakaaki

doi:10.1002/1097-0142(19830215)51:4<662::aid-cncr2820510419>3.0.co;2-x

Cited by 75 publications

(45 citation statements)

References 10 publications

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“…21 These cell lines were obtained from Cancer Research U.K. cell production services (London Research Institute Clare Hall Laboratories, Potters Bar, U.K.) and cultured in E4 medium supplemented with 10% heat-inactivated fetal calf serum (Gibco-Life Technologies, Paisley, U.K.) until harvest when they reached 80% confluence. Twenty-four hours before the cells were harvested for RNA isolation, the medium was replaced with E4 supplemented with 0.5% fetal calf serum.…”

Section: Cell Culture and Rna Isolationmentioning

confidence: 99%

Analysis of gene expression in cancer cell lines identifies candidate markers for pancreatic tumorigenesis and metastasis

Missiaglia

Blaveri

Terris

et al. 2004

Intl Journal of Cancer

139

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Pancreatic cancer is a highly aggressive type of malignancy and the prognosis for disease presenting typically at a late stage is extremely poor. A comprehensive understanding of its molecular genetics is required in order to develop new approaches to clinical management. To date, serial analysis of gene expression and more recently oligo/cDNA microarray technologies have been employed in order to identify genes involved in pancreatic neoplasia that can be developed as diagnostic markers and drug targets for this dismal disease. This study describes the expression profile obtained from 20 pancreatic cell lines using cDNA microarrays containing 9,932 human gene elements. Numerous genes were identified as being differentially expressed, some of which have been previously implicated in pancreatic adenocarcinoma (S100P, S100A4, prostate stem cell antigen, lipocalin 2, claudins 3 and 4, trefoil factors 1 and 2) as well as several novel genes. The differentially expressed genes identified are involved in a variety of cellular functions, including control of transcription, regulation of the cell cycle, proteolysis, cell adhesion and signaling. Validation of our array results was performed by exploring the SAGEmap database and by immunohistochemistry for a selection of 4 genes that have not previously been studied in pancreatic cancer: anterior gradient 2 homologue (Xenopus laevis), insulin-like growth factor binding protein 3 and 4 and Forkhead box J1. Immunostaining was performed using pancreas-specific tissue microarrays containing core biopsies from 305 clinical specimens. In addition, using statistical group comparison and hierarchical clustering, a selection of genes was identified that may be linked to the site of metastasis from which these cell lines were isolated.

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Section: Cell Culture and Rna Isolationmentioning

confidence: 99%

Analysis of gene expression in cancer cell lines identifies candidate markers for pancreatic tumorigenesis and metastasis

Missiaglia

Blaveri

Terris

et al. 2004

Intl Journal of Cancer

139

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“…The BxPc3 cell line (American Type of Culture Collection (ATCC), Rockville, Maryland) is a moderately to well differentiated cell line derived from a primary tumour (Tan et al, 1986) and expresses FGFR-1; the T3M4 cell line is a moderately differentiated cell line, which was derived from a lymph node metastasis (Okabe et al, 1983) and expresses FGFR-3; and the HPAF cell line which is a moderately to poorly differentiated cell line that was derived from ascitic fluid (Metzgar et al, 1982) and is FGFR-3 and FGFR-4 positive. All cell lines were maintained in standard medium.…”

Section: Cell Linesmentioning

confidence: 99%

FGF-1 and FGF-2 modulate the E-cadherin/catenin system in pancreatic adenocarcinoma cell lines

2001

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Summary Fibroblast growth factors (FGFs) and fibroblast growth factor receptors (FGFRs) have been increasingly recognized to play an important role in the pathobiology of pancreatic malignancy. We have investigated the effects of FGF-1 and FGF-2 on the behaviour and adhesion properties of human pancreatic adenocarcinoma cell lines (BxPc3, T3M4 and HPAF) that were previously characterised for the expression of FGFRs. Here we show that exposure to FGF-1 and FGF-2 leads to significant and dose-dependent increase in E-cadherindependent cell-cell adhesion, tubular differentiation, and a reduced capacity to invade collagen gels. FGF stimulation produces phosphorylation of E-cadherin and β-catenin on tyrosine residues, as well as increased E-cadherin localisation to the cytoplasmic membrane and association with FGFR1 demonstrable by coimmunoprecipitation. These results demonstrate that FGF-1 and FGF-2 may be involved in the regulation of cell adhesion, differentiation and invasion of pancreatic cancer. © Cancer Research Campaign http://www.bjcancer.com Growth factors and antibodiesHuman recombinant FGF-1 and FGF-2 (Upstate Biotechnology, Lake Placid, NY, USA) were used for stimulation of the cell lines at various concentrations between 1 ng/ml and 50 ng/ml. Heparin was added at 10 µg/ml and 1 µg/ml with FGF-1 and FGF-2, respectively, as shown to be suitable for FGFR activation by FGFs in this tissue type (Leung et al, 1994).The mouse monoclonal anti-human E-cadherin antibody, HECD-1, was kindly provided by Prof. Takeichi (Kyoto University, Kyoto, Japan). Mouse monoclonal anti-human E-cadherin, α-, β-and γ-catenin antibodies were purchased from Transduction Laboratories, Exeter, UK. The monoclonal anti-Ep-CAM (AUA-1) and anti-CEA (PR3B10) antibodies were kindly provided by Sir Walter Bodmer (ICRF). The anti-FGFR antibodies used in the study were anti-FGFR-1 antibodies (8E10 (Prizm Pharmaceuticals) and VBS6 (Santa Cruz)). Anti-phosphotyrosine antibody 4G10 was purchased from Upstate Biotechnology. Cell-cell adhesion assayCells were grown to 90% confluence, washed twice in PBS, and subsequently treated with 2 mM EDTA for 10 min at 37˚C. Detached cells were washed once with RPMI medium and were passed through Pasteur pipettes several times to obtain single cells. Cells were then re-suspended in either Ca 2+ -free PBS/0.8% FCS or RPMI/0.8% FCS (controls). In FGF stimulation experiments, FGF-1 or FGF-2 was added to the medium at concentrations of 1, 5, 10, 20, and 50ng/ml at the time of initiation of the assay. Cells were then inoculated at 5×10 5 cells/ml into 24-well plates (500µl/well) that had been coated overnight with 1% (w/v) bovine serum albumin in PBS to prevent non-specific cell adhesion to wells. Cells were allowed to aggregate for 1 h at 37˚C on a gyratory shaker with constant rotation at 90 rpm. Cell aggregation was terminated by the addition of 4% (w/v) glutaraldehyde fixative to individual wells at 0, 15, 30, 45 or 60 min. Aliquots were taken at each time point and the number of single cells was then determin...

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“…Human pancreatic adenocarcinoma cell lines AsPc-1 (Chen et al, 1982), BxPc-3 (Tan et al, 1986) and T3M-4 (Okabe et al, 1983) were obtained from Prof. N Lemoine (ICRF, London) and the American Tissue Culture Collection. All cells were maintained in RPMI-1640 culture medium with 10% fetal calf serum (FCS), 2 mM L-glutamine and antibiotics at 37°C in a humidified atmosphere containing 5% carbon dioxide.…”

Section: Cell Linesmentioning

confidence: 99%

Differential and antagonistic effects of 9-cis-retinoic acid and vitamin D analogues on pancreatic cancer cells in vitro

2000

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Retinoids and vitamin D are known to exert important anti-tumour effects in a variety of cell types. In this study the effects of 9- cis -retinoic acid (9cRA) the vitamin D analogues EB1089 and CB1093 on three pancreatic adenocarcinoma cell lines were investigated. All compounds caused inhibition of in vitro growth but the vitamin D analogues were generally the more potent growth inhibitors. They were also more effective on their own than in combination with 9cRA. Growth arrest correlated with an increased proportion of cells in the G0/G1 phase. Apoptosis was induced in the three cell lines by 9cRA, whereas neither EB1089 nor CB1093 had this effect. Furthermore, addition of EB1089 or CB1093 together with 9cRA resulted in significantly reduced apoptosis. Our results show that retinoic acids as well as vitamin D analogues have inhibitory effects on pancreatic tumour cells but different and antagonistic mechanisms seem to be employed. © 2000 Cancer Research Campaign

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Establishment and characterization of a carcinoembryonic antigen (CEA)-producing cell line from a human carcinoma of the exocrine pancreas

Cited by 75 publications

References 10 publications

Analysis of gene expression in cancer cell lines identifies candidate markers for pancreatic tumorigenesis and metastasis

Analysis of gene expression in cancer cell lines identifies candidate markers for pancreatic tumorigenesis and metastasis

FGF-1 and FGF-2 modulate the E-cadherin/catenin system in pancreatic adenocarcinoma cell lines

Differential and antagonistic effects of 9-cis-retinoic acid and vitamin D analogues on pancreatic cancer cells in vitro

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