摘要
目的:观察携带野生型pten基因的重组腺病毒(Ad-pten)体外转染高转移卵巢癌HO-8910PM细胞后的表达,并研究其对卵巢癌细胞抑制增殖、诱导凋亡和抑制迁移的作用.方法:利用Ad-Easy腺病毒载体系统构建Ad-pten,体外转染高转移卵巢癌HO-8910PM细胞,荧光显微镜检测Ad-pten的转染效率.Western印迹检测pten在HO-8910PM细胞中的表达;MTT实验观察pten对细胞生长的影响;HE染色法观察外源性pten基因表达对细胞形态学的影响;细胞划痕实验观察pten对细胞迁移的抑制作用;Hoechest33258凋亡染色检测pten基因对HO-8910PM细胞凋亡的诱导作用.结果:外源性野生型pten基因经腺病毒介导成功转入HO-8910PM细胞,Western印迹检测出有PTEN蛋白的表达,当感染复数MOI为100时,体外转染效率高达90%以上.pten显著抑制HO-8910PM细胞增殖、诱导其凋亡并能抑制其迁移.结论:重组腺病毒是高效的基因转移系统,能将pten目的基因高效地转移到高转移卵巢癌HO-8910PM细胞中,对细胞产生强有力的生长抑制及凋亡诱导作用,并能抑制其迁移.
AIM： To observe the expression of pten in human highly metastatic ovarian carcinoma cell line HO-8910PM after in vitro infection and investigate the effects of pten gene on the proliferation and metastatic ability of human ovarian carcinoma HO-8910PM cells. METHODS： Ad-pten was constructed through Ad-Easy adenovirus vector system and the expression of pten gene in HO-8910PM cells was determined by Western Blot. The transfection efficiency of Ad-pten was determined by fluorescence microscope. The inhibitive effect of Ad-pten on proliferation of HO-8910PM cells was determined by MTT assay. The morphologic changes of HO-8910PM cells transfected with Ad-pten were observed by light microscopy （ HE staining）. The metastatic ability was analyzed by cell scratching model in vitro. Hoechest33258 staining was used to study the cell apoptosis. RESULTS： The expression of PTEN protein in HO-8910PM cells was confirmed by Western Blot. The efficiency of adenovirus mediated Ad-pten gene transfer was above 90% when the multiplicities of infection （MOI） was 100. Exogenous pten gene significantly suppressed the growth and metastatic ability of HO-8910PM cells and induced the apoptosis of the cells. CONCLUSION： The constructed Ad-pten transfection system is highly efficient in transferring wild-type pten gene into human endometrial carcinoma cells. Ad-pten can strongly inhibit the proliferation and metastasis of the ceils, and also induce the apoptosis of the cells.
出处
《第四军医大学学报》
北大核心
2008年第3期231-234,共4页
Journal of the Fourth Military Medical University
基金
广东省自然科学基金(031962)
