RNA干扰肾细胞癌多药耐药基因1(MDR1)抑制癌细胞生长的机制探讨

Mechanism of Cell Growth Inhibition After the MDR1 Interfered by siRNA in Renal Cell Carcinoma

目的验证small interference RNA(siRNA)片段稳定转入肾细胞癌细胞中抑制其癌细胞生长,同时试图解释其原因。方法将合成好的RNAi慢病毒颗粒稳定转入目的细胞中并达到稳定传代后,以CCK-8试剂盒检测目的细胞的生长情况,以Real-time PCR检测目的细胞中p53基因及survivin基因的表达变化。结果 CCK-8法测得ACHN细胞在MDR1被干扰以后的生长水平,干扰组细胞明显低于两对照组,且干扰组细胞的细胞活力出现了明显的下降趋势。Real-time PCR检测得p53基因和survivin基因的mRNA表达水平明显降低,而无意义重组载体组(NC组)则和亲本细胞无明显差异。结论 MDR1沉默可使肾细胞癌ACHN细胞增殖减慢,生长抑制。同时沉默后细胞内的mtp53及survivin基因的表达均明显减少,MDR1介导的对肿瘤细胞生长的抑制作用至少部分是通过p53和survivin基因来实现的。

Objective To investigate the cell growth inhibition of the renal cell carcinoma following the small interference RNA fragment stabiling transfection,and explore the potential mechanism.Methods First,we transfered the RNA interference lentivirus particles into the target cells and made them stabling generating.Then we used the CCK - 8 kit to test the growth of the target cells and used the Real - time PCR to test the expression change of the P53 gene and the survivin gene.Results We find that the growth level of the interfered ACHN cell group was obviously lower than the other two control groups,and the cell vigor of the interfered group showed an obvious decrease.On the other hand,compared with the other two control groups,the mRNA expression level of the P53 gene and the survivin gene in the interfered group cells obviously depressed,but the NC group showed no much difference with the parent cells.Conclusions MDR1 gene silence can step down the cell proliferation of the renal cell carcinoma ACHN cells,and inhibit their growth.The expression of the mtP53 gene and survivin gene obviously deceased in the target cells when they were silenced.This demonstrated the MDR1 gene mediated inhibition would be implemented partially by the P53 gene and survivin gene.