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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

  • 摘要: 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-β E2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JEC 24h、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JEC 72h后,其OD值较对照组显著增高;(2)17-β E2(1×10-6mol/L)作用JEC细胞72h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-β E2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。

     

    Abstract: 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 1×10-7 and 1×10-6 mol/L concentrations of E2 were 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-β E2(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.

     

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