title: [Protective effect of lentivirus-mediated Bcl-2 gene transfection against phosphoramide mustard-induced apoptosis of human ovarian granulosa cells].
abstract: OBJECTIVE
  To investigate the effect of lentivirus-mediated Bcl-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 Bcl-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 Hochst 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±10.55)%, 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).
  
  
  CONCLUSIONS
  Lentivirus-mediated Bcl-2 gene transfection can protect human ovarian granulosa cells against PM-induced apoptosis by upregulating Bcl-2 protein expression.
authors: Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, China. douwangxuefeng@163.com
authors: Xuefeng
authors: X
authors: Wang
authors: Yuanli
authors: Y
authors: He
authors: Xiafei
authors: X
authors: Fu
authors: Dongxian
authors: D
authors: Peng
mesh_terms: D000328:Adult; D017209:Apoptosis / Q000187:drug effects; D005260:Female; D019254:Genes, bcl-2; D005822:Genetic Vectors; D006107:Granulosa Cells / Q000187:drug effects; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D010752:Phosphoramide Mustards; D014162:Transfection
chemical_list: D010752:Phosphoramide Mustards; C030090:phosphoramide mustard