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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 effects of different concentrations of estrogen (17-βE2) on the proliferation and cell cycle of estrogen receptor (ER) -negative endometrial adenocarcinoma cell line JEC. Methods The proliferative activity of human endometrial adenocarcinoma cell line JEC after addition of 17-βE2 was observed by cytometry, MTT assay and flow cytometry (FCM) Cell cycle phase changes were observed. Meanwhile, the expression of cyclin A in JEC cells was detected by immunohistochemistry and image analysis before and after the addition of 17-βE2. Results (1) The cell counts of JEC cells treated with 1 × 10-7 and 1 × 10-6mol / L estrogen for 4 days were significantly higher than that of the control group, while the other concentrations of E2 did not. After treated with different concentrations of E2 for 24 h and 48 h, the OD values measured by MTT method were not significantly different from those of the control group. The OD values of E2 treated with 1 × 10-6 mol / L of JEC for 72 h were significantly higher than those of the control group. (2) After 72h treatment with βE2 (1 × 10-6mol / L), the proportion of cells in S phase and G2 / M phase increased, and the proportion of cells in G0 / G1 phase decreased. However, the phase distribution of cell cycle did not change after 48h treatment. (3) The effect of 17-βE2 on the expression of cyclin A was significantly increased after JEC treatment. Conclusion Estrogen at a certain concentration can promote the proliferation of JEC cells in vitro with dose and time effect. Research suggests that this regulatory effect may be related to changes in cyclin A expression.