2.Effects of siRNA against Ki67 gene on the proliferation and apoptosis of human renal carcinoma cells
Jun-Nian ZHENG ; Qi-Duo SHI ; Xiao-Qing SUN ; Jia-Cun CHEN ; Ru-Min WEN ; Jing-Yi CAO ; Wen-Fa YANG ; Wang LI ; Jun-jie LIU ; Teng-xiang MA
Chinese Journal of Urology 2001;0(10):-
Objective To evaluate the effects of small interfering RNA(siRNA)against Ki67 gene on the proliferation and apoptosis of human renal carcinoma cell line 786-0 cells.Methods The human renal carcinoma 786-0 cells were treated with Ki67-siRNA(100 nmol/L).The mRNA expression of Ki67 was detected by RT-PCR.The protein expression of Ki67 was detected by Western blot and immunohisto- chemical technique,respectively.The proliferation of 786-0 cells was detected by MTT assay.The apoptosis of 786-0 cells was detected by TUNEL assay.Results RT-PCR and Western blot analysis showed that the Ki67 mRNA and Ki67 protein expression levels of the 786-0 cells treated with Ki67-siRNA were(37.6?1.9)% and(46.4?0.9)% ,respectively,which were significantly lower than those of controls [(97.3?0.9)% and(95.3?0.9)%,P<0.01],The Ki67 positive expression rate of 786-0 cells treated with Ki67-siRNA by immunohistochemical technique was 52.5?2.3,which was significantly lower than that of controls(114.5?4.9 ,P<0.01).The proliferation-inhibiting rate and apoptosis rate of the 786-0 cells trea- ted with Ki67-siRNA were( 63.6?1.6)% and(41.7?0.6)% ,respectively,which were significantly higher than those of controls [(2.8?0.2)% and(10.3?1.4)%,P<0.01].Conclusions siRNA against Ki67 gene can inhibit the proliferation and induce the apoptosis by blocking Ki67 expression of hu- man renal carcinoma 786-0 cells.The inhibition of Ki67 expression by siRNA may be a promising approach in gene therapy for renal cancer.