携带Bcl-2基因的慢病毒对磷酰胺氮芥抑制卵巢颗粒细胞体外分泌功能的影响

Lentivirus-mediated Bcl-2 gene expression in ovarian granulose cells improves cell secretion function

目的:探讨携带Bcl-2基因的慢病毒抑制磷酰胺氮芥(PM)对体外培养的人原代卵巢颗粒细胞(GCs)的凋亡情况及其分泌功能的影响。方法:实验分4组:实验组,pGC-FU-Bcl-2+PM;空载对照组,pGC-FU-EGFP+PM;实验对照组,只加PM;正常对照组,不加PM及病毒。采用DNA ladder检测GCs凋亡情况,利用放射免疫法检测细胞培养上清液中雌二醇及孕酮的浓度。结果:实验对照组及空载对照组的DNA ladder表现为明显的梯状带形,正常对照组及实验组DNA ladder无明显的梯状带形。各组之间雌二醇及孕酮的基础浓度经统计学分析表明差异无显著性,实验组与相应时间点实验对照组及空载对照组相比差异均存在显著性(P<0.01),实验对照组与空载对照组相比差异无显著性(P=0.990,P=0.523)。结论:携带Bcl-2基因的慢病毒感染人原代卵巢GCs后,显著降低了PM诱导的细胞凋亡,改善了GCs在体外分泌雌二醇及孕酮的功能。

Objective To investigate the effect of the lentivirus-mediated Bcl-2 gene expression on apoptosis and secretion function of the chemotherapy-injured human primary ovarian granulosa cells （GCs）. Methods Four groups were set in the present experiment： experimental group with pGC-FU-Bcl-2 ＋ PM, empty vector （EV） group with EV ＋ PM, experimental control group with PM, control group only with GCs culture. Apoptosis was detected by DNA-ladder assay. E2 and P in cell culture supernatant were determined by radiation immune assay. Results The obvious DNA ladder bands were shown in agarose gel electrophoresis in experimental control group and EV group, but not in the control group and experimental group. The concentration of E2 and P in experimental group was quite different on day 2 and 5 after adding the drug compared with that in the experimental group and EV group （P 〈 0.01）, but no significant difference in E2 and P concentration was found between the experimental group and EV group （P = 0.990, P = 0.523）. Conclusion The lentivirus-mediated Bcl-2 gene expression significantly reduced PM-induced apoptosis of GCs, which could improve secretion function of GCs in vitro.