Effects Of Silencing Kim-1Gene By SiRNA On Proliferation And Cell Cycle Of Renal Cell Carcinoma786-0Cells

ObjectiveTo investigate the effect of silencing Kidney Injury Molecular-1（Kim-1）geneby small interfering RNA（siRNA）interference on proliferation and cell cycle of renalcell carcinoma786-O cells. Explore Kim-1as a new therapeutic target of renal cellcarcinoma.MethodsThree diffierent siRNA(50nmol/L) which target human Kim-1gene wastransfected into786-O cells by lipofectamineTM2000. After24h,observe thetransfection efficiency by the fluorescence microscope. After48h,the expression ofKim-1was measured by quantitative real-time polymerase-chain-reaction（qRT-PCR）,choose the group which the silence effect is best for subsequent testing. After 24,48,72and96h,the proliferation active of each group’s cells was determined bymethylthiazoltetrazolium（MTT）assay respectively. After48h,the cell cycle of eachgroup’s cells was examined by flow cytometry.Results1. Liposome mediated transfection can effectively transfect siRNA into786-Ocells, the transfection efficiency is（94.36±2.57）%.2. In786-O cells the siRNA which targets Kim-1gene silenced the expressionof Kim-1signally. The silence effect of the three different Kim-1-siRNA sequences is（72.47±5.91）%、（12.98±2.21）%、（29.56±3.88）%respectively, however therelative expression quantity of the control group is（96.56±3.47）%（P<0.05）.3. In786-O cells after the silence of Kim-1gene, the proliferation of the cellis inhibited.96h after the transfection, the proliferation inhibition rate of the threegroup is0%(the blank control group),(3.80+1.24)%(the negative control group),(54.31+3.82)%(the experimental group) respectively（P<0.05）.4. In786-O cells the silence of Kim-1gene induced cell cycle arrest at G0/G1phases.48h after the transfection, the proportion of cells in G0/G1phase is (8.44±1.82)%(the blank control group),(42.06±2.15)%(the negative control group),(63.69±2.87)%(the experimental group) respectively（P<0.05）.Conclusion1. In renal cell cancer cells786-O, the Kim-1specific siRNA can effectivelysilence the expression of the Kim-1.2. In786-O cells, the down-regulation of Kim-1expression could induce theproliferation-inhibiting and arrest cell cycle at G0/G1phases.3. Kim-1molecule may play an important role in the occurrence and development of the renal cell carcinoma, and it can be used as a potential target of thespecific treatment in the renal cell carcinoma.