腺病毒载体介导的HPV16E7基因沉默及其对宫颈癌CaSki细胞的影响  

Adenovirus Vector Mediated HPV16E7 Silencing and its Effect on Cervical Cancer CaSki Cells

作　　者：李娜[1,2] 刘梦琼 郑义[1,2] 朱永强[1,2] 肖滑卫[1,2] 黄来强[1,2] 

机构地区：[1]清华大学生命科学学院,北京100084 [2]深圳市基因与抗体治疗重点实验室和生物医药研究中心.清华大学深圳研究生院生命与健康学部,广东深圳518055 [3]海南省人民医院,海南海口570311

出　　处：《现代生物医学进展》2013年第16期3001-3004,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(30671034);深圳市重点实验室提升计划;深圳市科技信息局基础研究计划(JC200903180532A)

摘　　要：目的:构建利用RNA干扰技术沉默HPV16E7基因的腺病毒载体,并探讨其对人宫颈癌细胞系CaSki细胞增殖的影响,以及腺病毒载体在宫颈癌基因治疗中的可行性。方法:利用腺病毒载体介导的RNA干扰对CaSki细胞中HPV16E7蛋白的表达进行抑制。显微镜观察细胞病变情况。MTT实验用来检测腺病毒载体感染的CaSki细胞的增殖情况。结果:成功构建了用以介导CaSki细胞中HPV16E7基因沉默的腺病毒载体。腺病毒感染后的CaSki细胞发生典型病理变化,HPV16E7基因表达受到明显抑制,细胞分裂明显减慢。结论:腺病毒载体介导的RNA干扰能够特异性的沉默HPV16E7基因,抑制CaSki细胞的增殖,为腺病毒载体用于宫颈癌基因治疗的可行性提供了依据。Objective： To construct an adenovirus vector for RNA interference of HPV16E7 gene,to investigate its effect on the proliferation of cervical cancer cell line CaSki,and to evaluate the availability of adenovirus vector as a media for cervical cancer gene therapy.Methods： The HPV16E7 gene was silenced by the adenovirus vector mediated RNA interference.The cytopathic effect of the infected CaSki cells was observed under microscope.The proliferation rates of CaSki cell line were measured by MTT assay.Results： Successfully constructed the adenovirus vector to inhibit the expression of HPV16E7 gene.Upon infection,the expression of HPV16E7 was reduced,typical cytopathic effect was observed,and the proliferation of CaSki cell was inhibited.Conclusions： The adenovirus mediated RNA interference can silence HPV16E7 expression,and can inhibit the proliferation of CaSki cell line,providing an evidence for the feasibility of adenovirus mediated gene therapy for cervical cancer.

关 键 词：宫颈癌 HPV16E7基因 腺病毒载体 RNA干扰 CASKI细胞 

分 类 号：R737.33[医药卫生—肿瘤] Q75[医药卫生—临床医学]