雌二醇对子宫内膜癌细胞MAPK通路的激活及其对增殖与迁移的影响

Activation of MAPK signaling pathway through estradiol and its effects on proliferation and migration in endometrial cancer cells

目的:探讨雌激素能否激活MAPK信号通路,及对子宫内膜癌细胞增殖能力的影响。方法:(1)不同浓度雌二醇作用于HEC-1A细胞后,蛋白印迹法检测p-ERK1/2蛋白活化水平。(2)实验分3组:E2组、U0126+E2组、对照组。荧光定量PCR技术检测细胞中MEK1/2、ERK1/2 m RNA的表达,蛋白印迹法检测p-MEK1/2、p-ERK1/2蛋白活化水平。(3)流式细胞仪,体外穿膜及细胞集落形成实验。结果:(1)不同浓度的E2作用细胞后,蛋白活化水明显高于对照组(P<0.05)。(2)蛋白活化水平E2组均高于对照组(P<0.05);U0126+E2组均低于E2组(P<0.05)。(3)G1期及S期细胞比例;穿过微孔的细胞数;集落形成率U0126+E2组与E2组,E2组与对照组比较,差异均有统计学意义(P<0.05)。结论 :雌二醇通过激活MAPK通路,促进子宫内膜癌的发生、发展。

Objective To explore whether estradiol can activate MAPK（mitogen activated protein kinase）pathway and its effect on the proliferation and migration in endometrial cancer cells. Methods There were 3groups including E2 group, U0126 ＋ E2 group and control group. p-ERK1 / 2 protein expression was detected by Western blot after stimulation of HEC-1A with different concentrations of estradiol. q RT-PCR and western blot were used to detect m RNA expression of MEK1 / 2 and ERK1 / 2 and protein of p-ERK1 / 2 and p-MEK1 / 2. Flow cytometry, transwell assay and colony formation were applied as well. Results The expression of p-ERK1 / 2protein was significantly higher in E2 group than that in control group（P 〈0.05）. The expressions of m RNA and protein of MEK1 / 2 and ERK1 / 2 were higher in E2 group than that in control group, but those in U0126 ＋ E2 group were lower than that in control group（P 〈0.05）. The percentage of HEC-1A in G1 phase and S phase, the cell number of migration and the colony forming rate in E2 group were higher than those in control group（P 〈0.05）, and those in U0126 ＋ E2 group were lower than those in E2 group（P 〈0.05）. Conclusion Estradiol can activate MAPK pathway, further promote the development of endometrial cancer cells.