摘要
目的研究刚地弓形虫RH株对体外培养的孕期小鼠胎盘滋养层细胞周期的影响及相关机制。方法制备小鼠胎盘滋养层细胞并分别接种于不同细胞培养皿,对照组加入DMEM高糖培养基孵育,实验组加入相同体积不同数量(2×104/mL、4×104/mL、8×104/mL)弓形虫速殖子培养6h、12h、24h;以MTT法检测滋养细胞增殖率;流式细胞仪检测细胞周期;Western印迹法检测细胞周期相关蛋白cyclinA、cdk2表达或活性。结果 MTT法检测结果显示刚地弓形虫RH株抑制滋养层细胞增殖,且呈时间剂量依赖性;流式细胞仪检测24h感染组细胞周期在S期产生阻滞,8×104/mL数量组S期细胞比例高达68.73%±1.76%;Western印迹方法分析感染弓形虫组滋养层细胞的cyclinA、cdk2蛋白表达量较对照组明显下降(P<0.05)。结论刚地弓形虫RH株能够抑制小鼠胎盘滋养层细胞增殖并通过调节cyclinA、cdk2等蛋白表达或活性改变引起细胞S期阻滞。
Objective To investigate the effects of Toxoplasma gondii RH strain on cell cycle of mouse placental tropho- blastic cells in vitro. Methods Mouse placental trophoblastic cells （ concentration of 5 x 104/mL） were harvested in the different cell culture plates. The cells were treated for different durations （6 - 24 h） with different concentrations of 7＂. gon- dii RH strain （ the concentration of tachyzoites was 2 x 104/mL, 4 x 104/mL, 8 x 104/mL, respectively）. Growth inhibition rates were measured with the MTT method ; Cell cycle was examined with flow cytometer. Western blot was used to detect the levels of cyclinA, cdk2 of ceils. Results T. gondii RH strain could inhibit the proliferation of trophoblastic cells in a time- dose dependent manner. Cell cycle was significantly stopped at GJM phase by T. gondii RH strain with FCM technology. Infection of T. gondii RH strain reduced the expressions of gene cyclinA and cdk2 of trophoblastic cells. Conclusion In- fection of T. gondii RH strain could inhibit trophoblastic cells and arrest the cell cycle mainly by regulating the expression of gene cyclinA and cdk2.
出处
《四川动物》
CSCD
北大核心
2012年第6期900-904,共5页
Sichuan Journal of Zoology
关键词
刚地弓形虫RH株
滋养细胞
细胞周期
Toxoplasma gondii RH strain
trophoblastic cells
cell cycle
