Two new human cholangiocarcinoma cell lines and their cytogenetics and responses to growth factors, hormones, cytokines or immunologic effector cells

Abstract

Two new human cholangiocarcinoma (CC) cell lines (CC-SW-I and CC-LP-I) were established and maintained in culture for 2 years. Histologically, both original liver tumors were adenocarcinomas, and the cell lines exhibited morphologic features of moderately differentiated adenocarcinoma. Immunohistochemistry showed that both cell lines were strongly positive for cytokeratin AEI but negative for carbohydrate tumor-associated antigen, CA19-9. Ultrastructural analysis of both cell lines showed the presence of tight junctional complexes and focally formed microvilli. Both CC cell lines were tumorigenic in nude mice. Cytogenetic analysis showed that both cell lines expressed highly aneuploid karyotypes with numerous structural and numerical deviations. CC-SW-I was hypodiploid with numerous chromosome losses and structural rearrangements, while CC-LP-I was hyperdiploid and displayed multiple additional chromosomes. Doubling times for the CC-SW-I and CC-LP-I cell lines in the presence of 15% fetal bovine serum were 72 hr and 180 hr, respectively. Growth of the CC-SW-I cell line was significantly stimulated in the presence of insulin, while that of the CC-LP-I cell line was significantly augmented by epidermal growth factor (EGF). In contrast, dexamethasone strongly inhibited proliferation of both cell lines in a dose-dependent manner. Among various recombinant cytokines examined for effects on growth or surface antigen expression on CC cell lines, only interleukin I-beta (ILI-beta) strongly inhibited growth of the CC-LP-I cell line, while interferons (IFNs) or tumor necrosis factor-alpha (TNF-alpha) were mildly inhibitory. Both tumor cell lines were resistant to natural killer (NK) cells but sensitive to lymphokine-activated killer (LAK) cells. Preincubation of tumor cells with IFN-gamma, IFN-alpha or TNF-alpha significantly decreased the susceptibility of each tumor cell line to lysis by LAK cells, and the change in sensitivity did not correlate with the expression of HLA antigens or intercellular adhesion molecule-I (ICAM-I) on the surface of tumor cells. These 2 CC cell lines are expected to provide valuable information about cell biology of human CC.

Involvement of 85-kd cytosolic phospholipase A2 and cyclooxygenase-2 in the proliferation of human cholangiocarcinoma cells

“…3 This study was designed to determine whether there is an association between cytokine-or growth factor-mediated cholangiocarcinoma cell growth and PG metabolism in the human cholangiocarcinoma cell lines SG231, 33 HuCCT1, and CC-LP-1. 34 We report that both HGF and IL-6 induced rapid phosphorylation of cPLA 2 and release of AA and PG from SG231 cells. Addition of PGE 2 and PGF 2 ␣, alone, to SG231 cells results in significantly increased cell proliferation.…”

“…Three human cholangiocarcinoma cell lines, SG231(33), CC-LP-1 (34), and HuCCT1 (obtained from Japanese Health Science Research Resources Bank), were utilized in this study. SG231 cells were cultured in serum-supplemented medium (minimum essential medium with 2 mmol/L Lglutamine, 50 g/mL gentamicin, 10 mmol/L HEPES, and 10% fetal bovine serum).…”

“…1,[4][5][6][7] In this study, experiments were designed to investigate whether AA metabolism is involved in HGFand IL-6-induced proliferation of human cholangiocarcinoma cell lines SG231, 33 HuCCT1, and CC-LP-1. 34 The effects of specific inhibitors of PLA 2 and COX-2 in HGFand IL-6-induced cholangiocarcinoma cell growth were also investigated. Although it might be difficult to exclude completely the potential nonspecific effect of chemical inhibitors in human cells, utilization of established cPLA 2 and COX-2 inhibitors in this study would provide evidence for the involvement of AA metabolism in HGFand IL-6-induced cholangiocarcinoma growth.…”

PPARγ ligands inhibit cholangiocarcinoma cell growth through p53-dependent GADD45 and p21WAF1/Cip1 pathway

“…Three human cholangiocarcinoma cell lines, including SG231, 36 CC-LP-1, 37 and HuCCT1 (obtained from Japanese Health Science Research Resources Bank, Osaka, Japan) were used in this study. SG231 cells were cultured in serum-supplemented medium (SSM) (␣-minimum essential medium with 2 mmol/L L-glutamine, 50 g/mL gentamicin, 10 mmol/L HEPES, and 10% fetal bovine serum).…”

AdEasy-based cloning system to generate tropism expanded replicating adenoviruses expressing transgenes late in the viral life cycle

“…To demonstrate that fiberlinked transgene expression depends on E1 expression, E1-positive cell lines (HEK293, HER911, GH329) and E1-negative cell lines (HeLa and CC-LP-1) were transduced with AdFiber-GFP and analyzed for DsRed and eGFP expression by flow cytometry. [21][22][23][24] E1-negative cells were transduced with higher multiplicities of infection (MOIs) and analyzed at later time points (at least 3 days post-transduction) to ensure that low levels of background expression could be detected. Still as expected and summarized in Table 1, no detectable eGFP expression was observed in E1-negative cells while both single-(DsRed) and double-(DsRed and eGFP) positive cells were detected in E1-positive cells.…”