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。 |
关键词: 肝肿瘤 胆管上皮细胞 HepG2细胞 共培养 caspase3 细胞凋亡 |
DOI:10.3724/SP.J.1008.2011.017 |
投稿时间:2010-09-14修订日期:2010-10-28 |
基金项目:广东省自然科学基金(4009383). |
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Biliary epithelial cells inhibit proliferation of co-cultured hepatic cancer cell line HepG2 |
HE Yao1*,HUANG Wen-sheng2,YANG Rong-ping1,REN Ming1 |
(1. Department of Gastroenterology, the First Affiliated Hospital of SUN Yat-sen University, Guangzhou 510080, Guangdong, China;;2. Department of Surgery, the First Affiliated Hospital of SUN Yat-sen University, Guangzhou 510080, Guangdong, China) |
Abstract: |
ObjectiveTo investigate the possible effect of mouse intrahepatic biliary epithelial cell line (mIBEC) on cocultured human hepatoma cell lines HepG2.MethodsHepG2 and mIBEC cells were cocultured in a membraneseparated 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 cocultured mIBEC. Realtime PCR(RTPCR)was used to determine Ki67 and caspase3 mRNA expression in HepG2 cells in a system with or without cocultured mIBEC. Western blotting analysis was used to determine caspase3 protein level in HepG2 cells. ResultsThe proliferation of the cocultured HepG2 cells was significantly lower than those cultured alone(P<0.01). Expression of Ki67,a cell proliferation marker,was also significantly downregulated in mIBEC cocultured HepG2 cells (P<0.05). The levels of caspase3 mRNA and protein were significantly upregulated in mIBEC cocultured HepG2 cells compared with HepG2 cells cultured alone (P<0.05). ConclusionmIBEC can inhibit the proliferation of cocultured HepG2, and caspase3 activation might be one of the reasons for the inhibitory effect of mIBEC against HepG2 cells. |
Key words: liver neoplasms biliary epithelial cells HepG2 cells co-culture caspase3 apoptosis |