The Study Of The Inhibition Of Human Renal Cancer Cell Line 786-O By Silencing Survivin Gene With RNA Interference

Background and Objective:Renal cell carcinoma is one of the most common malignant tumor in the urinary system. In China, the incidence of renal cell carcinoma ranks the 2nd in the urinary tumor. The renal clear cell carcinoma(RCCC) is the most common pathological type of renal cell carcinoma, accounts for about 70%～80%. As the early symptoms was not obvious, the majority of patients are at the advanced stage at the time of diagnosis, and because of the majority of the renal cell carcinoma are not sensitive to chemotherapy and radiotherapy, most of the advanced renal cell carcinoma patients lack of effective therapy . So It's very important and necessary to find a practical and effective treatment. RNAi is the breaking of a double-stranded RNA (dsRNA) matching a specific gene sequence into short pieces called short interfering RNA, which trigger the degradation of mRNA that matches its sequence. As a one of the gene blocking technologies, RNAi has been a research hotspot in recent years and rapidly developed,and it is expected to become a new important tool of anti-tumor. As a member of the inhibitor of apoptosis protein (IAP) family discovered in recent years,survivin has been taken more and more attention in the role of tumor's occurrence and development ,and it's expected to become a new target for cancer gene therapy. In this experiment,on the basis of detect the survivin gene express significantly in human renal cell carcinoma 786-O cells , we design and synthesis siRNA that match of the survivin gene, transfect it into the cells to block the survivin gene's expression in human renal cancer cell line 786-O specifically,and to observe the inhibitory effects of RNAi inducing the apoptosis of human renal cancer cell line 786-O.Methods:(1)The growth curve of renal carcinoma cell line 786-O were detected by MTT,the expression of survivin mRNA and protein in human renal cancer cell line 786-O were detected by RT-PCR and immunohistochemisty.(2) The target gene fragments were cloned into pcDNA?6.2-GW/EmGFPmiR vectors , and then the recombinant eukaryotic expression vectors were transfected into 786-O cells with liposomes.The expression changes in survivin mRNA and protein were detected by RT-PCR and immunohistochemisty,the proliferation inhibitory rate (IR) of 786-O cells was detected by MTT.Results:(1) The survivin mRNA and protein were found in human renal cancer cell line 786-O.In contrast,no expression of survivin was found in normal renal cells. (2) The mRNA and protein expressions of survivin gene in the siRNA group were significantly inhibited by the eukaryotic expression vectors , and it also inhibit significantly the growth and proliferation of 786-O cells.Conclusion : The survivin gene and protein were significantly expressed in renal clear cell carcinoma 786-O. Silencing survivin gene by the RNAi technique can not only decrease effectively the expressions of survivin gene and protein,but also inhibit the cell proliferation in 786-O cells.And this may suggested that the survivin gene play an important role in the renal cancer's occurrence and development, it may provide the experimental evidence for the survivin gene as a target of gene therapy in renal cancer.