Characterization of a New Primary Human Pancreatic Tumor Line

Mh, Tan; Nj, Nowak; Loor, Rueyming; Ochi, Hisako; Aa, Sandberg; Lopez, Charles D.; Jw, Pickren; Berjian, R; Ho, Douglass; Tm, Chu

doi:10.3109/07357908609039823

Cited by 201 publications

(129 citation statements)

References 16 publications

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“…Pancreatic carcinoma cell lines AsPC-1 (ATCC, CRL-1682), BxPC-3 (Tan et al, 1986), MiaPaCa-2 (ATCC, CRL-1420), Panc-1 (ATCC, CRL-1469), PaTu8988t (differentiated) and PaTu8988s (undifferentiated, metastases forming) (these two lines are from the same patient (Elsasser et al, 1993)) were cultured in Dulbecco's Modified Eagle Medium with 10% FCS; Capan-1 (ATCC, HTB-79), SW850 and SW-979 in Click's RPMI Medium with 10% FCS. Cells were seeded in tissue culture dishes and cultured at 378C for 3 -5 days, harvested in sub-confluent state and lysed as described below.…”

Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

Analysis of the cytochrome c-dependent apoptosis apparatus in cells from human pancreatic carcinoma

2002

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Defects in the apoptotic system are likely to play a role in tumorigenesis. Pancreatic carcinoma cells are extremely resistant to apoptosis induction by chemotherapy suggesting that the apoptosis machinery is faulty. We investigated the integrity of the cytochrome c-dependent apoptotic apparatus in 10 human pancreatic carcinoma cell lines. Expression of Apaf-1, caspase-3, -6, -7, -8 and -9, Hsp-70 and XIAP was detected in all cell lines. The expression levels of Apaf-1 and caspase-8 were homogenous in all cell lines whereas differences in expression of other caspases were seen. In cytosolic fractions, all investigated caspases were processed in response to cytochrome c but the extent of processing varied between the cell lines. No stringent correlation between the amount of processing of caspase-9 and effector caspases was seen. Cytochrome cinduced effector caspase activity was quantitated by enzyme assay. Especially at low concentrations of added cytochrome c, this response varied greatly between the cell lines. These data demonstrate that the apoptotic system downstream of the mitochondria is qualitatively intact in pancreatic carcinoma. They further show that the response to cytochrome c can be quantitated in a cell-free system and that determinants other than mere expression of apoptotic molecules can regulate cytochrome c-induced apoptosis.

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Section: Cell Lines and Culture Conditionsmentioning

confidence: 99%

Analysis of the cytochrome c-dependent apoptosis apparatus in cells from human pancreatic carcinoma

2002

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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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Characterization of a New Primary Human Pancreatic Tumor Line

Cited by 201 publications

References 16 publications

Analysis of the cytochrome c-dependent apoptosis apparatus in cells from human pancreatic carcinoma

Analysis of the cytochrome c-dependent apoptosis apparatus in cells from human pancreatic carcinoma

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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