siRNA抑制鼻咽癌HNE-1细胞株HPV16 E6的表达和细胞增殖

siRNA inhibits HPV16 E6 expression and cell proliferation in HNE-1 cell line

目的:探讨小干扰RNA(Small interferirng RNA,siRNA)对鼻咽癌(Nasopharyngeal carcinoma,NPC)HNE-1细胞株HPV16E6基因表达的抑制作用,观察HPV16E6基因沉默对鼻咽癌细胞生长的影响。方法:体外合成4条针对HPV16E6基因的siRNA,通过脂质体转染导入鼻咽癌HNE-1细胞。用MTT检测转染后癌细胞的生长增殖情况,用半定量逆转录聚合酶链反应(Reverse transcription polymerase chain reaction,RT-PCR)检测转染后HPV16 E6 mRNA的表达水平,用免疫组化技术检测鼻咽癌细胞中HPV16 E6蛋白的表达水平。结果:导入有效siRNA后,MTT实验显示细胞的生长增殖受到明显抑制,抑制率可达32%;PT-PCR及免疫组化结果显示RNA干扰(RNA interfering,RNAi)可使HNE-1细胞中HPV16 E6 mRNA水平及蛋白表达水平下降。结论:siRNA可以有效抑制HNE-1细胞株中HPV16 E6的表达,降低鼻咽癌细胞的增殖能力。RNAi技术作为一种新的基因治疗技术在鼻咽癌治疗方面有着巨大的应用潜力。

Objective：To evaluate the inhibitory effect of Small interferirng RNA（siRNA） on human papilloma vinus（HPV）16 E6 oncogenes in nasopharyngeal carcinoma（NPC） cell line HNE-1,and observe the effects of HPV16 E6 silencing on NPC cell growth. Methods：Four siRNA against the HPV16 E6 gene were designed and transfected into HNE-1 cell respectively using RNAi-Mate transfection reagent. MTT was used to investigate the cellular proliferation after the transfection. RT-PCR was used to detect the expression of HPV16 E6 and protein level of HPV16 E6 was measured by Immunohistochemical staining. Results：After the most effective siRNA was transfected into HNE-lcell,the result of MTT indicated that the cellular proliferation was restrained remarkably,and the highest inhibitory, rate was 32%. Meanwhile,RT-PCR and immunohistochemical staining showed that there was a significant decrease in HPV16 E6 mRNA and protein level. Conclusion：These results demonstrated that siRNA against HPV16 E6 could effectively downregulate HPV16 E6 expression in HNE-1 cell line,and inhibit the cellular proliferation. RNAi has great potential in the treatment of nasopharyngeal carcinoma as a new gene therapy.