凋膜止崩液对人离体子宫内膜增生症腺上皮细胞PTEN、Rb2/p130表达影响

Impacts of Diaomo Zhibeng Fluid on PTEN and Rb2/p130 Expression in Epithelial Cells of Human Endometrial Hyperplasia

目的：通过凋膜止崩液对人离体子宫内膜增生症（EH）腺上皮细胞PTEN、Rb2/p130基因表达的影响,探讨其对EH的治疗机理。方法：分离、培养30例原代人子宫内膜增生症（EH）腺上皮细胞并采用CK-19鉴定,将纯化后的腺上皮细胞随机分为4组：空白对照组、凋膜止崩液大剂量组（50μg/mL）、凋膜止崩液中剂量组（25μg/mL）、凋膜止崩液小剂量组（12.5μg/mL）,分别于用药24、48、72 h观察下列指标的变化：四甲基偶氮噻唑兰（MTT）法检测3个时间点各组细胞的增殖率、实时定量聚合酶连反应（RT-PCR）以及Western blot分别检测各组细胞PTEN、Rb2/p130 m RNA及蛋白表达的差异。结果：与空白对照组比较,凋膜止崩液各组腺上皮细胞的增殖率明显降低,且呈时间和剂量依赖性（P〈0.05）;凋膜止崩液组EH腺上皮细胞中PTEN、Rb2/p130 m RNA及蛋白的表达明显增加,与空白对照组相比差异显著（P〈0.05）。结论：凋膜止崩液能明显抑制EH腺上皮细胞的增殖,其抑制作用与上调细胞抑癌基因PTEN、Rb2/p130的表达有关。

Objective：To reveal the underlying mechanism of Diaomo Zhibeng Fluid（DZF）in treating endometrial hyperplasia（EH）by investigating the impacts of DZF on PTEN and Rb2/p130 expression in isolated epithelial cells of human EH.Methods：Primary glandular epithelial cells（GPCs）from 30 cases EH patients were isolated,cultured,and then identified by CK-19.The purified GPCs were randomly divided into four groups：blank control group with no DZF,DZF large dose group（50 μg/mL）,DZF middle dose group（25 μg/mL）,DZF small dose group（12.5 μg/mL）.After giving DZF for 24 h,48 h or 72 h,the following indexes were examined in each time point：Thiazolyl Blue Tetrazolium Bromide（MTT）method was used to detect cell proliferation rates,Real-time PCR and Western blot were used to detect the differences in m RNAs and proteins of PTEN and Rb2/p130 between DZF groups and control group.Results：Compared with blank control group,GPC proliferation rate was significantly reduced in each DZF treated groups（P0.05）,in a time and dose dependent manner.In addition,the m RNAs and proteins of PTEN and Rb2/p130 were significantly increased（P0.05）in DZF treated groups.Conclusion：DZF can significantly inhibit the proliferation of EH cells,which is probably achieved through the up-regulation of tumor suppressor genes PTEN and Rb2/p130.