南方医科大学学报2012，Vol.32Issue(7)：932-936,5.DOI:10.3969/j.issn.1673-4254.2012.07.005
慢病毒介导的Bcl-2基因对磷酰胺氮芥诱导人原代卵巢颗粒细胞凋亡的保护作用
Protective effect of lentivirus-mediated Bcl-2 gene transfection against phosphoramide mustard-induced apoptosis of human ovarian granulosa cells
摘要
Abstract
Objective To investigate the effect of lentivirus-mediated Bd-2 gene transfection in protecting human primary ovarian granulosa cells against phosphoramide mustard (PM)-induced apoptosis. Methods Granulosa cells were isolated from the follicle fluid of women undergoing in vitro fertilization and embryo transfer. The lentiviral vectors carrying Bd-2 gene (pGC-FU-Bcl-2) and enhanced green fluorescence protein (pGC-FU-EGFP) were constructed and packaged into high-titer lentiviruses. The resulting recombinant lentivirus carrying Bcl-2 and EGFP genes or the empty vector were used to infect the primary human ovarian granulosa cells, followed by addition of PM in the cell culture, with untreated granulosa cells as the control. The cell apoptosis was detected by Annexin V and Horchst 33258 staining, and the expression of Bcl-2 protein was assessed using Western blotting. Results The control granulosa cells showed an apoptotic rate of (1.93+0.28)%. The cells infected with pGC-FU-Bcl-2 prior to PM exposure had a apoptotic rate of (6.99±0.55)%, significantly higher than that of the control cells, but significantly lower than that of the cells with PM exposure only and those infected with the empty vector before PM exposure (P<0.05). The expression of Bcl-2 was the highest in the cells infected with pGC-FU-Bcl-2 prior to PM exposure (P<0.05). Conclusion Lentivirus-mediated Bcl-2 gene transfection can protect human ovarian granulosa cells against PM-induced apoptosis by upregulating Bcl-2 protein expression.关键词
Bcl-2基因/慢病毒/化疗药物/细胞凋亡/原代卵巢颗粒细胞Key words
apoptosis/ chemotherapeuhc agents/ ovarian granulose cells/ Bcl-2 gene/ lentivirus vector分类
医药卫生引用本文复制引用
王雪峰,何援利,付霞霏,彭冬先..慢病毒介导的Bcl-2基因对磷酰胺氮芥诱导人原代卵巢颗粒细胞凋亡的保护作用[J].南方医科大学学报,2012,32(7):932-936,5.基金项目
国家自然科学基金(81041101) （81041101）
广东省自然科学基金(10451051501004704) （10451051501004704）
