DNA-PKcs影响肝癌耐药细胞BEL7402/5-FU耐药性的机制

目的利用RNA干扰技术探讨DNA-PKcs基因沉默影响肝癌细胞BEl7402/5-FU耐药性的机制。方法将靶向DNA-PKcs的干扰质粒shDNA-PKcs转染肝癌耐药细胞BEL7402/5-FU,利用实时荧光定量PCR及Western blot法检测转染干扰质粒后DNA-PKcs基因的沉默效率;Western blot检测Artemis、磷酸化Artemis蛋白水平的表达。结果实时荧光定量PCR及Western blot检测结果显示转染shDNA-PKcs后,DNA-PKcs mRNA及蛋白水平表达均下调（P〈0.05）;Western blot检测结果显示shDNA-PKcs转染组Artemis蛋白表达水平与对照组相比均有所降低（P〈0.05）,磷酸化Artemis蛋白表达水平在转染前后无明显变化（P〉0.05）。结论 ShDNA-PKs干扰质粒能抑制Bel-7402/5FU细胞中DNAPKcs、Artemis蛋白的表达、但对P-Artemis的表达无影响。

The effect of DNA-PKcs gene silence on the expression of Artemis and P-Artemis in liver cancer BEL7402 /5-FU cells

Objective To observe the effect of DNA-PKcs gene silencing on the mechanism of drug resistance in Bel7402 /5-FU by RNA interference technology.Methods Targeted DNA-PKcs interference plasmid was transfected into liver cancer cell BEL7402 /5-FU.DNA-PKcs silencing efficiency was detected by Real-time PCR and Western blot.The expression of Artemis and P-Artemis by Western blot.Results Real-time PCR and Western blot assay indicated that the mRNA and protein levels of DNA-PKcs expression were downregulated after transfection of shDNA-PKcs（ P〈0.05）.Western blot analysis showed that the expression of Artemis was decreased on protein levels compared with the control group（ P〈0.05）;P-Artemis protein expression level was not changed before and after transfection compared with the control group（ P〉0.05）.Conclusion The shDNA-PKcs reduces DNA-PKcs and Artemis expression in BEL7402 /5-FU cells,but had no effect on the expression of P-Artemis.