siRNA介导的HPV16E6/E7基因沉默对宫颈癌SiHa增殖及凋亡的影响  

Effects of siRNA-mediated HPV16 E6 / E7 transcriptional gene stable silencing on proliferation and apoptosis of human cervical cancer SiHa cells

作　　者：王平[1] 刘珊[2] 程波[2] 吴西钊[2] 武迪 许琳[2] 石建玲[2] 段恋[2] 孙锁柱[2] 

机构地区：[1]河北北方学院病理学与病理生理学系,河北张家口075000 [2]第二炮兵总医院病理科,北京100088 [3]大兴区人民医院病理科,北京102600

出　　处：《诊断病理学杂志》2015年第12期781-786,共6页Chinese Journal of Diagnostic Pathology

摘　　要：目的探讨小干扰RNA(siRNA)介导的人乳头状瘤病毒16型(HPV16)E6/E7基因沉默对宫颈癌Si Ha细胞增殖及凋亡的影响。方法设计靶向HPV16 E6/E7基因启动子区的siRNA序列,构建质粒转染Si Ha细胞,筛选稳定沉默E6/E7基因的细胞系。用QT-PCR和RT-PCR检测细胞E6/E7基因mRNA的变化、Western blot检测细胞E6/E7蛋白变化;利用MTT、RT-PCR、免疫组化法检测细胞增殖情况,Tunel检测细胞的凋亡情况。结果成功构建E6/E7基因稳定沉默的细胞株,其mRNA与蛋白沉降效率明显。MTT及Ki-67表达显示细胞增殖活性降低;Tunel显示沉默细胞凋亡增加。结论 siRNA技术可以成功诱导HPV16型E6/E7基因沉默,从而影响癌细胞的生长、增殖和凋亡。Objectives To investigate human papillomavirus（ HPV） type 16 E6 / E7 gene silencing induced by promotertargeting siRNA and its effect on proliferation and apoptosis of human cervical cancer Si Ha cells.Methods In this study,we designed siRNA targeting promoter of human papillomavirus 16 E6 / E7 and transfected it by plasmid into the cervical cancer cell line,Si Ha.E6 / E7 mRNA and protein expression were respectively detected by quantitative time-polymerase chain reaction（ QT-PCR） and reverse transcription-polymerase chain reaction（ RT-PCR） and Western blot.Futhermore,cellular growth and proliferation were detected by MTT,QT-PCR and immunohistochemistry（ IHC）.Cellular apoptosis were detected by TUNEL.Results We had successfu L established E6 / E7 gene silencing stable cell line named cm-16 cells,its mRNA and protein were simultaneously reduced.The proliferation activity of cm-16 cells was significantly lower than Si Ha,however,the apoptosis was significantly higher than Si Ha.Conclusion siRNA effectively knocks down both extraneous HPV E6 / E7 gene,and then effects cellular growth,proliferation and apoptosis.

关 键 词：SIRNA HPV 细胞增殖与凋亡 SIHA细胞 

分 类 号：R73-35[医药卫生—肿瘤]