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

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.