RNA干涉宫颈癌细胞抑制HPV16 E6基因表达对FHIT基因表达的影响

Effect of inhibited HPV16 E6 expression by RNA interference on FHIT gene expression in cervical cancer cells

目的:研究HPV16 E6基因被RNA干涉抑制后FHIT基因表达的变化,探讨两者在宫颈癌组织中的相关性。方法:合成针对HPV16 E6基因的特异性si RNA,脂质体介导转染宫颈癌CaSki细胞,RT-PCR测定转染前后HPV16E6、FHIT基因mRNA表达;Western blot和流式细胞仪检测转染前后HPV16E6、FHIT蛋白表达情况。结果:将HPV16 E6 si RNA转染CaSki细胞后48h,HPV16E6mRNA表达水平较转染前降低了80.3%,P<0.01;FHIT mRNA的表达水平较转染前升高了41.7%,P<0.01。转染前后HPV16E6蛋白表达水平分别为26.7±3.3和8.2±1.1,t=14.224,P=0.005;FHIT蛋白表达水平分别为73.5±5.6和109.9±5.1,t=22.018,P=0.002。结论:抑制HPV16 E6基因的表达可使宫颈癌组织中FHIT基因的表达增加。

OBJECTIVE：To detect the changes of FHIT gene expression after the HPV16 E6 gene expression was inhibited by RNA interference, and explore the relationship between HPV16 E6 and FHIT gene in cervical neoplasms. METHODS：The specific small interfering RNA （siRNA） of HPV16 E6 gene was synthesized and transfected into cervical cancer cell line CaSki. The mRNA levels of HPV16 E6 and FHIT before and after transfection were measured by RT PCR, and the protein levels of HPV16 E6 and FHIT were measured by Western blot and flow cytometry. RESULTS： The mRNA level of HPV16 E6 reduced by 80.3% at 48 hours after transfection （P〈0.01）, but the mRNA level of FHIT was 41.7 % higher than before at 48 hours after transfection （P〈0.01）. The protein levels of HPV16 E6 before and after transfection were （26.7±3.3） and （8.2± 1.1）, respectively,t 14.224,P=0.005; The protein levels of FHIT before and after transfection were （73.5±5.6） and （109.9±5.1）,respectively,t= 22. 018,P=0. 002. CONCLUSION： FHIT gene expression can be up regulated by inhibition of the expression of HPV16 E6 gene in cervical neoplasms.