RNA干扰HPV E6下调eIF4E转录表达抑制宫颈癌Hela细胞增殖并影响细胞周期进程研究.docVIP

RNA干扰HPV E6下调eIF4E转录表达抑制宫颈癌Hela细胞增殖并影响细胞周期进程研究 　　[摘要] 目的 探讨宫颈癌Hella细胞中干扰HPV E6对eIF4E表达的调控，探寻HPV E6促癌的新途径，为宫颈癌诊断和治疗提供新靶点。 方法 构建高效的靶向HPV18 E6的shRNA质粒（shE6）并测序，而后转染人宫颈癌Hela细胞，荧光显微镜及流式细胞术检测转染效率。而后以Real-time PCR检测E6及eIF4E的mRNA水平，采用CCK-8法检测Hela细胞增殖能力的改变，流式细胞术检测细胞周期。 结果 测序结果显示成功构建shE6 质粒。shE6-3转染人宫颈癌Hela细胞后48 h，E6及eIF4E mRNA表达的抑制率分别约为80%，76%。shE6-3对Hela细胞的增殖活性在48 h的抑制效果最明显，增殖抑制率约为21%；相对于NC组，shE6组G0/G1期细胞比例显著增高，而S期比例减少，G2/M期未见明显变化。 结论 RNA干扰HPV E6能够下调eIF4E转录表达，抑制宫颈癌Hela细胞增殖，并阻滞细胞周期于G0/G1期。eIF4E有望成为宫颈癌防治的潜在靶点。 　　[关键词] 宫颈癌；E6；真核细胞翻译起始因子；RNA干扰 　　[中图分类号] R737.33 [文献标识码] A [文章编号] 1673-7210（2014）01（b）-0004-04 　　Study of RNA interference of HPV E6 down regulates the transcripiton of eIF4E and inhibits the cell proliferation and cell cycle of Hela cells 　　WANG Sen1 GAO Min2 ZHAO Yi1 ZHU Wei1 JIANG Enping1 KANG Haixian1 YAO Yunhong1 HU Xinrong1 　　1.Department of Cancer Institute and Pathology， Guangdong Medical College， Guangdong Province， Guangzhou 523808， China； 2.Department of Pathology， the People′s Hospital of Dongguan， Guangdong Province， Dongguan 523059， China 　　[Abstract] Objective To investigate RNA interference of E6 could regulate eIF4E in cervical cancer cells and to find new target gene for the diagnosis and therapy of cervical cancer. Methods ShRNAs of E6 were constructed by company and transfected into Hela cells. The transfection rate was detected by fluorescence microscope and flowcytometry （FCM）. Real time PCR was used to detected mRNA of E6 and eIF4E. CCK-8 was employed for cell proliferation and FCM was used to detected the cell cycle. Results ShRNA plasmids of E6 were constructed successfully and transfected into Hela cells. 48 h later， the mRNA of E6 and eIF4E were 80%， 76%， respectively. ShE6-3 leaded to the inhibition of cell proliferation （21%） at 48 h. In the cell cyle detection， the cells of G0/G1 phase increased highly while the cells of S phase decreased. The cells of G2/M phase was the same to that of the control group. Conclusion ShE6 can inh