HBx不同表达水平对HepG2肝癌细胞增殖、侵袭以及索拉非尼耐药的影响  

The effects of HBx on the proliferation and invasion of hepatocellular carcinoma and sorafenib resistance

作　　者：刘路政 陈家诚 陈良 陈骋 许达峰 林仕勋 罗相相 武金才 Liu Luzheng;Chen Jiacheng;Chen Liang;Chen Cheng;Xu Dafeng;Lin Shixun;Luo Xiangxiang;Wu Jincai(Department of Hepatobiliary and Pancreatic Surgery,Hainan Affiliated Hospital of Hainan Medical University(Hainan General Hospital),Haikou 570311,China)

机构地区：[1]海南医学院附属海南医院(海南省人民医院)肝胆胰外科,海口570311

出　　处：《中华肝胆外科杂志》2021年第11期842-846,共5页Chinese Journal of Hepatobiliary Surgery

基　　金：国家自然科学基金(81660489);海南省社发重点项目(ZDYF2020134);海南省自然科学基金(819QN356);海南省研究生创新科研项目(Hys2020-355)。

摘　　要：目的探讨HepG2肝癌细胞不同乙型肝炎病毒X(HBx)表达水平对增殖、侵袭以及索拉非尼耐药的影响。方法肝癌HepG2细胞株感染HBx高表达序列的慢病毒为HBx高表达组,阴性对照组同HBx高表达组,但仅感染空载体。干扰组在HBx高表达组的基础上感染HBx干扰序列病毒,降低HBx表达。干扰对照组同干扰组,但感染的空载体。空白对照组无任何处理。Western印迹检测HBx的蛋白表达。细胞增殖检测试剂盒检测增殖,Transwell侵袭实验检测侵袭能力,检测索拉非尼半抑制浓度(IC50)。结果Western印迹结果显示成功构建各组稳定表达细胞株。HBx高表达组细胞增殖能力优于空白对照组、阴性对照组,干扰组细胞增殖能力低于干扰对照组和HBx高表达组,差异均有统计学意义(均P<0.05)。空白对照组、阴性对照组、干扰组穿过Matrigel胶的细胞数分别为(46.2±4.1)个、(50.7±5.1)个、(48.2±5.2)个。HBx高表达组穿过Matrigel胶的细胞数(124.2±8.3)个、干扰对照组(117.2±7.5)个,均高于上述三组,差异均有统计学意义(均P<0.05)。HBx高表达组和干扰对照组细胞IC50分别为(5.36±0.31)μmol/L和(5.48±0.20)μmol/L,高于空白对照组、阴性对照组、干扰组的(4.75±0.22)μmol/L、(4.60±0.14)μmol/L和(3.98±0.03)μmol/L,差异均有统计学意义(均P<0.05)。结论HBx高表达促进HepG2肝癌细胞增殖和侵袭,提高HepG2肝癌细胞对索拉非尼的抵抗能力,抑制细胞凋亡。Objective To investigate the effects of hepatitis B virus X(HBx)on hepatocellular carcinoma(HCC)proliferation,invasion,and sorafenib resistance.Methods HepG2 cell line infected with HBx ORF lentivirus was set as the HBx high expression group and infected with empty vector was set as the negative control group.The interference group was infected with the HBx siRNA virus based on the HBx high expression group to reduce HBx expression.Interference control group as interference group but with infected empty vector virus.Western blotting was used to measure the protein level of HBx.Cell proliferation,invasion ability,and sorafenib semi-inhibitory concentration(IC50)of HCC cells under different HBx expression levels were respectively detected by cell proliferation assay kit,Transwell invasion assay,and cell titer-glo kit.Results Western blotting showed that the stable cell lines were successfully established.Cell proliferation of the HBx high expression group was better than that of the blank control and negative control groups,and the cell proliferation of the interference group was lower than that of the interference control and HBx high expression groups,and the differences were all statistically significant(P<0.05).The number of cells crossing Matrigel gel was(46.2±4.1),(50.7±5.1)and(48.2±5.2)in the blank control group,negative control group,and interference group,respectively.The number of cells crossing Matrigel gel in the HBx high expression group(124.2±8.3)and the interference control group(117.2±7.5)were higher than the above three groups,respectively,and the differences were all statistically significant(P<0.05).The IC50 of cells in the HBx high expression group and the interference control group were(5.36±0.31)μmol/L and(5.48±0.20)μmol/L,respectively,which were higher than those in the blank control group,the negative control group,and the interference group(4.75±0.22)μmol/L,(4.60±0.14)μmol/L and(3.98±0.03)μmol/L.The differences were all statistically significant(P<0.05).Conclusion HBx promote

关 键 词：癌 肝细胞 乙型肝炎病毒X 索拉非尼 靶向耐药 

分 类 号：R735.7[医药卫生—肿瘤]