2-ME抑制人子宫内膜癌细胞株增殖的研究

Study on inhibition of 2-methoxyestradiol on proliferation in endometrial carcinoma cells

目的 探讨2-甲氧雌二醇(2-ME)对人子宫内膜癌细胞株KLE细胞体外增殖和凋亡的抑制作用.方法选用人子宫内膜癌细胞株KLE进行体外培养,实验组加入不同浓度2-ME的培养液,对照组不含2-ME.用四甲基偶氮唑蓝(MTT)比色法观察2-ME对人子宫内膜癌细胞株KLE增殖的抑制作用;药物作用后的克隆形成实验;电子显微镜(电镜)观察细胞形态变化;流式细胞仪(FCM)观察细胞的凋亡率及细胞周期的变化.结果2-ME浓度为10.0～50.0μM时,明显抑制KLE细胞的增殖(P<0.01),并具有时间依赖性和剂量依赖性.2-ME作用后G0/G1期细胞增加,并伴随G0/G1期细胞的增加,出现细胞凋亡峰和凋亡率的升高(P<0.05).电镜下观察到KLE细胞染色体边集、核固缩、凋亡小体.结论2-ME对人子宫内膜癌KLE细胞株增殖有抑制作用,并能促进其凋亡.

Objective To investigate the inhibition of 2-methoxyestradiol (2-ME) on proliferation and apoptosis in human endometrial cell line KLE in vitro. Method Endometrial cancer cell line KLE originated from human endometrial adenocarcinoma was cultured in vitro. Study group (2-ME in different concentrations.) and control group without 2-ME . Cell proliferation was measured by 3-(4,5-dimethylthiazol-z-yl)-2,5-dipheny tertrazolium blue (MTT) colorimetric assay; Cell cycle and apoptotic percentage were detected by flow cytometry (FCM); morphological changes of apoptotic cells were observed by electron microscopy; Clone forming test was used to determine the proliferating capability after 2-ME treatment. Results 2-ME inhibited KLE cells growth significantly in a does-dependent and time-dependent manner(P<0.01). After treatment with 2-ME ,the enhanced G_0/G_1 arrest was accompanied with the enhanced apoptotic peak and percentage, as well apoptotic cells were found more than those in control group(P<0.05). By electron microscopy, there were many morphological characteristics of apoptosis including compaction and margination of nuclear chromatin, nuclear fragments and apoptotic bodies. Conclusions It is suggested in the present study that 2-ME can inhibit the proliferation of KLE cells in vitro and induce them apoptosis.