miR-26b对肝癌HepG2奥沙利铂耐药细胞的增敏作用研究

Sensitization of miR-26b to oxaliplatin-resistant hepatocellular carcinoma HepG2 cells

目的:构建肝癌HepG2奥沙利铂耐药细胞,观察miR-26b在其耐药机制中的作用。方法:使用浓度梯度法构建肝癌奥沙利铂耐药细胞株(HepG2/L-OHP)。miR-26b mimic转染HepG2/L-OHP细胞,CCK-8法检测细胞增殖,Real-time PCR定量检测miR-26b表达,Western blot检测蛋白表达。结果:HepG2细胞miR-26b的2-△△Ct值为0.206±0.024,显著高于HepG2/L-OHP细胞的0.152±0.017(P<0.01)。奥沙利铂对HepG2、HepG2/L-OHP和转染miR-26b mimic的HepG2/L-OHP细胞的IC50值分别为22.15±3.26、48.56±6.47和36.73±5.18μmol/L,各组间差异有统计学意义(P<0.01)。HepG2、HepG2/L-OHP和转染miR-26b mimic的HepG2/L-OHP细胞的LC3-Ⅱ/Ⅰ比值为2.27±0.24、9.45±1.54和6.32±1.13,各组间差异有统计学意义(P<0.01)。结论:miR-26b可以抑制肝癌奥沙利铂耐药细胞自噬激活,逆转肝癌细胞对奥沙利铂耐药。

Objective: To establish oxaliplatin-resistant hepatocellular carcinoma cell line(HepG2/L-OHP) and observe the change of sensitivity of oxaliplatin in miR-26b-overexpression HepG2/L-OHP cells. Methods: The HepG2/L-OHP cell line was established by concentration gradient method. HepG2/L-OHP cells were transfected by miR-26b mimic. mi R-26 b was detected by Real-time PCR.The cell proliferation was completed by CCK-8 analysis. The protein expression was determined by western blot. Results: 2-△△Ctvalues of miR-26b in HepG2 cellswas 0.206 ±0.024, which was significantly higher than that of 0.152±0.017 in HepG2/L-OHP cells(P<0.01). The IC50 values of oxaliplatin to HepG2, HepG2/L-OHP, and mi R-12 b transfected HepG2/L-OHP cells was 22.15 ±3.26, 48.56 ±6.47, and 36.73 ±5.18μmol/L. There were significant differences among the three groups(P<0.01). The value of LC3-Ⅱ/Ⅰ in HepG2, HepG2/L-OHP, and miR-12b transfected HepG2/L-OHP cells was 2.27 ±0.24, 9.45 ±1.54, and 6.32±1.13. There were significant differences among the three groups(P<0.01). Conclusion: miR-26b can inhibit the autophagic activation of oxaliplatin-resistant hepatocellular carcinoma cells and reverse the oxaliplatin-resistant in hepatocellular carcinoma.