胆管上皮细胞株抑制共培养的肝癌HepG2细胞株增殖
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广东省自然科学基金(4009383).


Biliary epithelial cells inhibit proliferation of co-cultured hepatic cancer cell line HepG2
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Supported by Natural Science Foundation of Guangdong Province(4009383).

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    摘要:

    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 96R○ 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。

    Abstract:

    ObjectiveTo investigate the possible effect of mouse intrahepatic biliary epithelial cell line (mIBEC) on cocultured human hepatoma cell lines HepG2.MethodsHepG2 and mIBEC cells were cocultured in a membraneseparated Transwell system. CellTiter 96R○ AQueous One Solution Cell Proliferation Assay (Promega Corporation,Madison,WI) was used to examine HepG2 cell proliferation in the system with or without cocultured mIBEC. Realtime PCR(RTPCR)was used to determine Ki67 and caspase3 mRNA expression in HepG2 cells in a system with or without cocultured mIBEC. Western blotting analysis was used to determine caspase3 protein level in HepG2 cells. ResultsThe proliferation of the cocultured HepG2 cells was significantly lower than those cultured alone(P<0.01). Expression of Ki67,a cell proliferation marker,was also significantly downregulated in mIBEC cocultured HepG2 cells (P<0.05). The levels of caspase3 mRNA and protein were significantly upregulated in mIBEC cocultured HepG2 cells compared with HepG2 cells cultured alone (P<0.05). ConclusionmIBEC can inhibit the proliferation of cocultured HepG2, and caspase3 activation might be one of the reasons for the inhibitory effect of mIBEC against HepG2 cells.

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  • 收稿日期:2010-09-14
  • 最后修改日期:2010-10-28
  • 录用日期:2010-12-09
  • 在线发布日期: 2011-01-20
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