目的:探讨孕激素是否通过其受体调控乙二醛酶Ⅰ(GloⅠ)的表达,进而调控子宫内膜癌细胞的增殖和凋亡活性。方法:应用Westen blot检测孕激素对GloⅠ,Caspase3,Cyclin D1表达的影响;经孕激素受体(PR)特异性抑制剂RU-486处理后,用Westen blot检测PR介导的GloⅠ,Caspase3,Cyclin D1分子的表达;应用siRNA干扰技术敲除GloⅠ表达后,检测GloⅠ对caspase3和Cyclin D1表达的影响,并分别应用MTT和TUNEL检测孕激素通过GloⅠ介导对细胞增殖和凋亡的影响。结果:(1)孕激素呈剂量依赖效应地下调GloⅠ、Cyclin D1的表达、上调Caspase3的表达,与对照组相比差异有统计学意义(P〈0.05);(2)MPA+RU-486组GloⅠ,Cyclin D1的蛋白水平比RU-486组、MPA处理组增加,Caspase3的表达下降,差异有统计学意义(P〈0.05);(3)siGloⅠ干扰组GloⅠ蛋白表达水平比阴性对照组明显降低,干扰效率达50%(P〈0.05),GloⅠ被干扰后,加入孕激素细胞的增殖活性明显受到抑制,凋亡活性增加,差异有统计学意义(P〈0.05)。结论:孕激素可通过PR调控GloⅠ介导的子宫内膜癌细胞增殖和凋亡活性。
Objective:To investigate whether MPA regulates the expession of GloⅠ through the progesterone receptor,following regulates endometrial cancer cell proliferation and apoptotic activity.Methods:Westen blot was used to detect the expression of GloⅠ,Caspase3,Cyclin D1 in protein levels after treatment with the progesterone;After the treatment of PR specific inhibitor,Westen blot was used to detect the expression of GloⅠ,Caspase3,Cyclin D;siRNA was used to konck down the expression of GloⅠ gene and MTT assay was used to determine cellular growth,TUNEL was used to determine cellular apoptosis.Results:(1)The expression of GloⅠ,Cyclin D1 could be down-regulated by MPA in Ishikawa cells and the expression of Caspase3 could be up-regulated by MPA in dose-dependent(P0.05).(2)The expression of GloⅠ,Cyclin D1 was up-regulated and the Caspase3 was down-regulated by MPA±RU-486 compared with control group(P0.05).3.Inhibition of GloⅠ by RNA interference could reduce cell proliferation and increase cell apoptosis(P0.05).Conclusion:MPA can regulate cell proliferation and apoptosis activity by GloⅠ gene through the progesterone receptor.