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pten基因重组腺病毒对卵巢癌细胞生长的抑制作用

Inhibition on growth of ovarian carcinoma cells by recombinant adenovirus carrying pten gene
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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.
作者 林观平 熊亮 李树梅 黄秀兰 周克元
机构地区 广东医学院生物化学与分子生物学教研室 赣南医学院预防医学教研室 攀枝花钢铁(集团)公司第十九冶职工医院妇产科
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)
关键词 卵巢肿瘤 基因治疗 PTEN基因 腺病毒 细胞凋亡 迁移抑制 ovarian neoplasms gene therapy pten gene adenovirus apoptosis inhibition of metastasis
分类号 R733.7 [医药卫生—肿瘤]
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参考文献9

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二级参考文献21

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第四军医大学学报
2008年 第3期

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