重组腺病毒介导的RbAp48基因表达对人宫颈癌细胞生长、增殖的影响  

Effects of enhanced expression of RbAp48 mediated by recombinant adenovirus on proliferation and growth of cervical cancer cells

作　　者：张雯[1] 冯婷婷[1] 郑琳[1] 王红[1] 卢翌[2] 齐眉[1] 于修平[1] 唐伟[1] 赵蔚明[1] 

机构地区：[1]山东大学医学院医学微生物学教研室,山东济南250012 [2]山东大学医学院生物化学与分子生物学系,山东济南250012

出　　处：《山东大学学报（医学版）》2014年第4期13-17,共5页Journal of Shandong University:Health Sciences

基　　金：国家自然科学基金(81101980;81272209);山东省自然科学基金(ZR2010HQ001;ZR2012HM031)

摘　　要：目的构建携带RbAp48基因序列的重组腺病毒,检测其对宫颈癌细胞系SiHa肿瘤细胞行为的影响。方法构建穿梭质粒pDC316-RbAp48,与骨架质粒pBHGloxΔE1,3Cre共转染HEK293细胞,获得复制缺陷型重组腺病毒Ad5-RbAp48,Ad5-RbAp48感染人宫颈癌细胞SiHa,采用免疫印迹法检测RbAp48表达,使用WST-8、细胞计数及软琼脂克隆形成实验,研究Ad5-RbAp48介导RbAp48高表达对SiHa细胞增殖与非锚定依赖性生长的影响。结果成功构建并鉴定重组腺病毒Ad5-RbAp48,Ad5-RbAp48感染SiHa细胞导致RbAp48高效表达,并有效抑制SiHa细胞增殖能力与非锚定依赖性生长能力。结论 RbAp48重组腺病毒可介导RbAp48高效表达,具有抑制宫颈癌细胞肿瘤细胞行为的作用。Objective To construct the recombinant adenovirus carrying RbAp48 gene, and to study its effect on the be-haviors of human cervical cancer cell line SiHa.Methods The shuttle plasmid pDC316-RbAp48 was constructed and co-transfected into the HEK293 cells with genomic plasmids pBHGloxΔE1, 3Cre to produce the replication-deficient re-combinant adenovirus Ad5-RbAp48.Then the SiHa cells were infected with Ad5-RbAp48, and RbAp48 expression was examined by immunoblotting.WST-8, cell number counting, and soft agar assay were carried out to examine the effects of Ad5-RbAp48 infection on cell proliferation and anchorage-independent growth of SiHa cells.Results Ad5-RbAp48 was successfully constructed.Ad5-RbAp48 infection significantly enhanced the expression of RbAp48 in SiHa cells, and effectively inhibited cell proliferation and anchorage-independent growth.Conclusion Recombinant adeno-virus carrying RbAp48 gene can mediate RbAp48 expression and inhibit behaviors of cervical cancer cells.

关 键 词：宫颈癌 RbAp48 腺病毒 基因表达 

分 类 号：R37[医药卫生—病原生物学]