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目的通过体外共培养肝内胆管上皮细胞株(mIBEC)与肝癌细胞株HepG2,探讨mIBEC对HepG2细胞株的可能作用。方法体外共培养HepG2与mIBEC,采用CellTiter 96○R AQueous One Solution Cell Proliferation Assay分别测定并比较HepG2细胞单独培养时以及与mIBEC共培养后增殖的情况;实时定量PCR法分别测定HepG2细胞单独培养时以及与mIBEC共培养后其Ki67及caspase3 mRNA表达水平;蛋白质免疫印迹法分别测定HepG2细胞单独培养时以及与mIBEC共培养后其caspase3蛋白表达水平。结果与mIBEC共培养后24～72 h,HepG2细胞增殖水平低于其单独培养时的水平(P<0.01),且伴有Ki67 mRNA表达的下调(P<0.05);与mIBEC共培养后,HepG2细胞caspase3 mRNA及蛋白表达水平较其单独培养时均上调,差异有统计学意义(P<0.05)。结论 mIBEC可抑制共培养的HepG2细胞增殖;caspase3激活表达上调可能是mIBEC抑制共培养的HepG2细胞增殖的机制之一。
Objective To explore the possible effect of mIBEC on HepG2 cell line by co-culturing intrahepatic cholangiocarcinoma cell line (mIBEC) and hepatoma cell line HepG2 in vitro. Methods HepG2 and mIBEC were co-cultured in vitro. The cell proliferation and proliferation of HepG2 cells were detected by CellTiter 96 ○ R AQueous One Solution Cell Proliferation Assay. The proliferation of HepG2 cells was evaluated by real-time quantitative PCR. After co-culture with mIBEC, the expression of Ki67 and caspase3 mRNA were detected by Western blotting. The expression of caspase 3 protein in HepG2 cells cultured alone and co-cultured with mIBEC were detected by Western blot. Results The proliferation of HepG2 cells at 24-72 h after co-culture with mIBEC was lower than that of HepG2 alone (P <0.01), and the expression of Ki67 mRNA was down-regulated (P <0.05). After co-cultured with mIBEC, HepG2 The expression of caspase3 mRNA and protein were up-regulated when compared with those cultured alone. The difference was statistically significant (P <0.05). Conclusion mIBEC can inhibit the proliferation of HepG2 cells. The up-regulation of caspase 3 activation may be one of the mechanisms by which mIBEC inhibits the proliferation of HepG2 cells.