转染HBx诱导肝癌细胞株Bel-7404多药耐药的初步研究

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Abstract Objective To study the effects of HBx transfection on proliferation, cell cycle, and multidrug resistance of hepatocellular carcinoma cell line Bel-7404, and to investigate the possible mechanism of HBx transfection in inducing the multidrug resistance of liver cancer. Methods Human hepatocellular carcinoma cell line Bel-7404 was transfected, and the cells were divided into three groups according to the transfection conditions, which were transfected HBx cell group (Bel-7404-HBx group), transfected cell group with empty vector (Bel-7404-con group) and blank cells group (Bel-7404 group). The expression of HBx RNA in all groups was detected by real-time PCR. The expression of HBx, Notch-1 and multi-drug resistance protein (MRP) in all groups was detected by Western blot, the cell proliferation activity and multidrug resistance in all groups were determined by CCK-8, and the cell growth cycle was analyzed by flow cytometer. Results PCR amplified gel electrophoresis showed that HBx fragments were detected in Bel-7404-HBx group, while no HBx mRNA expression could be found in the other two groups; Western blot indicated that the protein band (HBx protein) was detected in the Bel-7404-HBx group, while no expression was found in Bel-7404 group and Bel-7404-con group. Furthermore, compared with Bel-7404 group and Bel-7404-con group, the cell proliferation in Bel-7404-HBx group was increased significantly (P ＜ 0.05); compared with Bel-7404 group and Bel-7404-con group, the protein expression of Notch-1 and MRP in Bel-7404-HBx group was increased significantly; the results of cell cycle showed that, compared with Bel-7404 group and Bel-7404-con group, the number of cells in G0/G1 phase was decreased significantly (P ＜ 0.05), which was significantly increased in S phase (P ＜ 0.01), while there was no significant differences in G2/M phase (P ＞ 0.05); multidrug resistance experiments showed that, compared with Bel-7404 group and Bel-7404-con group, the resistance index of Mitomycin, Adriamycin, 5-Fluorouracil, Cisplatin and Oxaliplatin in the Bel-7404-HBx group was increased significantly (P ＜ 0.05 or P ＜ 0.01). Conclusion Bel-7404 cells transfected by HBx may induce overexpression of Notch-1 and MRP, leading to the increase of multiplication capacity of hepatocellular carcinoma cell line Bel-7404 and the occurrence of multidrug resistance.

Key words： HBx Hepatocellular carcinoma cell line Bel-7404 Notch signaling pathway Multi-drug resistance protein Multidrug resistance

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	LIN Songting CHEN Zhengyi ZHONG Wenzhou

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LIN Songting CHEN Zhengyi ZHONG Wenzhou. Primary study on HBx transfection in inducing the multidrug resistance of hepatocellular carcinoma cell line Bel-7404[J]. 中国医药导报, 2017, 14(27): 16-20.

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http://www.yiyaodaobao.com.cn/EN/ OR http://www.yiyaodaobao.com.cn/EN/Y2017/V14/I27/16

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