雌激素对ER阴性的子宫内膜腺癌细胞JEC增殖的作用

Effect of Estrogen on Growth of Estrogen Receptor Negative Endometrial Adenocarcinoma Cell Line JEC in vitro

目的研究不同浓度的雌激素(17-βE2)对雌激素受体(ER)阴性的子宫内膜腺癌细胞系JEC增殖和细胞周期的影响。方法采用细胞计数法、四甲基偶氮唑蓝比色法(MTT)和流式细胞术(FCM)的方法,观察人子宫内膜腺癌细胞系JEC在加入17-βE2后的增殖活性和细胞周期时相变化;同时用免疫组化及图像分析,检测JEC细胞在加入17-βE2前后细胞周期调控蛋白cyclin A表达的变化。结果(1)1×10-7和1×10-6mol/L的雌激素作用JEC细胞4天后,细胞计数均明显高于对照组,而其余浓度E2无明显作用。不同浓度E2作用JEC24h、48h后,MTT法测OD值和对照组相比无明显差异;1×10-6mol/L的E2作用JEC72h后,其OD值较对照组显著增高;(2)17-βE2(1×10-6mol/L)作用JEC细胞72h后使S期及G2/M期的细胞比例增加,G0/G1期细胞比例减少。但作用48h后细胞周期时相分布无明显变化;(3)17-βE2作用JEC后可使细胞内cyclin A蛋白表达明显增加。结论一定浓度的雌激素能促进JEC细胞体外增殖,且具有剂量和时间效应性。研究提示,这种调控作用可能与cyclin A表达的变化有关。

Objective To study the effect of different concentration of estrogen（E2） on the growth of estrogen receptor negative of endometrial adenocarcinoma cell line JEC in vitro. Methods Endometrial adenocarcinoma cell line JEC originated from human endometrial adenocarcinoma was cultured in vitro. Two groups were set up： sample group （17-β estradiol at different concentrations） and control group without estrogen. The proliferative capacity of endometrial adenocarcinoma cell line JEC in the culture medium with 17-β estradiol was assessed by cell counting on a haemocytometer and evaluated by the means of 3- （4,5-dimethylthiazol z yl）-2,5-dipheny tertrazolium blue （MTT）and cell cycle was analyzed by flow cytometry （FCM）. The expression of cyclin A of JEC was examined by immunocytochemical staining and using automatic image analysis technology. Results （1）The cell numbers in the fourth day after dealing with 1×10^-7and 1×10^-6 mol/L concentrations of E2were more than that in control group. The optical density of JEC cell in the 72th hour after dealing with 1 ×10^-6 mol/L concentration of E2 was higher than that in control group（P〈0. 01）. There was no statistics significant difference in the other experimental groups compared with the control group（P〉0. 05）. （2）FCM showed that 17-13 F4 （1×10^-6 mol/L） increased the cells percentage at the S and G2/M phases of cell cycle and decreased the cells percentage in G0/G1 phases of cell cycle. （3）Analysis on expression of intracellular cyclin A protein showed that estradiol could obviously up-regulate cyclin A protein by the immunohistochemical method of SABC. Conclusion It is suggested in the present study that estrogen can stimulate the proliferation of endometrial adenocarcinoma cell line JEC in vitro. There is an accordant dose-response and time-response relationship. In addition, its effect may be associated with the change in the expression of cyclin A.