FXYD6过表达对肝细胞癌SMMC7721细胞5-Fu化疗敏感性的影响

Effect of FXYD6 overexpression on tchemosensitivity of hepatocellular carcinoma SMMC7721 cells treated with 5-Fu

目的观察FXYD离子转运调节因子6(FXYD6)过表达对肝细胞癌SMMC7721细胞5-氟尿嘧啶(5-Fu)化疗敏感性的影响。方法将肝细胞癌SMMC7721细胞分为过表达组和对照组,分别转染pcDNA3.1/FXYD6和pcD-NA3.1/Vector真核质粒,培养48 h筛选SMMC7721抗性克隆。取两组转染后的抗性克隆细胞,分别加入终浓度为0、12.5、25、50、100μg/mL的5-Fu作用24、36、48 h,采用MTT法检测细胞增殖抑制率。取两组转染后的细胞,加入终浓度为25μg/L的5-Fu作用48 h,采用流式细胞术检测细胞凋亡率。结果随着5-Fu浓度增加和作用时间延长,两组细胞增殖抑制率均呈升高趋势。5-Fu浓度为12.5μg/mL时,两组作用24、36、48 h的细胞增殖抑制率比较均无明显差异(P均>0.05);5-Fu浓度为25、50、100μg/mL时,过表达组作用24、36、48 h的细胞增殖抑制率均明显低于对照组同时间点(P均<0.01)。观察组细胞凋亡率为35.62%±3.98%,对照组为46.19%±6.60%,两组比较P<0.01。结论 FXYD6蛋白过表达可降低肝细胞癌SMMC7721细胞对5-Fu的化疗敏感性。

Objective To investigate the effect of FXYD6 overexpression on the chemosensitivity of hepatocellular carcinoma SMMC7721 cells treated with 5-fluorouracil(5-Fu).Methods SMMC7721 cells were randomly divided into the overexpression group and control group.The SMMC7721 cells were transfected with pcDNA3.1/FXYD6 and pcD⁃NA3.1/Vector eukaryotic plasmids respectively,cultured for 48 h to screen the SMMC7721 resistant clones.Then pcD⁃NA3.1/Vector and pcDNA3.1/FXYD6 transfected SMMC7721 clones were obtained.When the two groups were added with final concentrations of 5-Fu at 0,12.5,25,50 and 100μg/mL after 24,36,and 48 h respectively,the cell prolifer⁃ation inhibition rate was detected by MTT method.When the two groups were added with final concentration of 5-Fu(25μg/L)for 48 h,the apoptosis rate was detected by flow cytometry.Results With increase of 5-Fu concentration and time extension,the cell proliferation inhibition rates of the two groups showed an increasing trend.When 5-Fu was 12.5μg/mL,there was no significant difference in the inhibition rate of cell proliferation between the two groups at 24,36,and 48 h(all P>0.05).When 5-Fu was 25,50,and 100μg/mL,the cell proliferation inhibition rates of the overexpression group at 24,36,and 48 h were significantly lower than those of the control group at the same time point(all P<0.01).The apoptosis rate of the observation group was lower than that of the control group(35.62%±3.98%vs.46.19%±6.60%,P<0.01).Conclusion Overexpression of FXYD6 can reduce the chemosensitivity of hepatocellular carcinoma SMMC7721 cells treated with 5-Fu.