解放军医学杂志2012，Vol.37Issue(8)：760-764,5.
携带Bcl-2基因的慢病毒对原代培养的人卵巢颗粒细胞的影响
Effect of lentiviral-mediated Bcl-2 gene on primary human ovarian granulosa cells
摘要
Abstract
Objective To investigate the infectious effect of lentivirus carrying Bcl-2 gene on primary human ovarian granulosa cells (GCs) and its potentiality to induce apoptosis. Methods The lentivirus vector carrying Bcl-2 gene (pGC-FU-EGFP-bcl-2) was constructed and condensed to a high titer. The recombinant lentivirus with different MOI (10, 50, 100, 200, 400) were used to infect primary human ovarian GCs, and then the infection rate and cell proliferation of GCs were observed at 24, 48, 72 and 96 hours after infection. The GCs were incubated for 24 hours, and then divided into 3 groups: experimental group (GC-FU-Bcl-2 with MOI 100 was added into GCs), normal control group (only GCs), and empty vector control group (only the empty vector pGC-FU-EGFP was added into GCs). The expressions of Bcl-2 protein and mRNA in GCs were detected by Western blotting and RT-PCR, respectively, at the 3rd and 7th day after infection, and the apoptosis rate of GCs at day 3 was detected by flow cytometry Results The primary human ovarian GCs began to adhere with colony formation after 24h growth, and they were polygonal or spindle-shaped. When MOI was 100, the GCs maintained normal morphology and growth, and the infection rate in GCs reached the peak value (60%) after 72h cultivation. After being infected by the lentivirus carrying Bcl-2 gene, the expressions of Bcl-2 mRNA and protein could be detected in the GCs of experimental group, and the apoptotic rate of GCs was reduced significantly compared with that in empty vector control group (P<0.05)> Conclusion It seems that the lentivirus carrying Bcl-2 gene could increase the expression of Bcl-2 protein and, accordingly, inhibit the apoptosis of primary ovarian GCs.关键词
基因,bcl-2/慢病毒感染/卵巢/粒层细胞/细胞凋亡Key words
genes, bcl-2/ lentivirus infections/ ovary/ granulosa cells/ apoptosis分类
医药卫生引用本文复制引用
王雪峰,何援利,刘木彪,付霞霏..携带Bcl-2基因的慢病毒对原代培养的人卵巢颗粒细胞的影响[J].解放军医学杂志,2012,37(8):760-764,5.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）
