雌激素通过钙蛋白酶上调P21/CCNE2 mRNA表达并促进乳腺癌细胞MCF-7的侵袭能力  

Estrogen up-regulates expressions of P21 and cyclin E2 mRNA and enhances MCF-7 cell invasion via activation of calpain

作　　者：王丹丹[1] 王筑婷[2] 杨莉[1] 宛蕾[3] 王旭东[1] 

机构地区：[1]贵州医科大学基础医学院生理学教研室,贵州贵阳550025 [2]贵州医科大学基础医学院生物化学教研室,贵州贵阳550025 [3]贵州医科大学基础医学院药理学教研室,贵州贵阳550025

出　　处：《基础医学与临床》2016年第12期1636-1640,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(31360252)

摘　　要：目的探讨17β-雌二醇(E2)对乳腺癌细胞mRNA表达,观察E2侵袭的分子信号机制。方法以乳腺癌细胞MCF-7为观察模型,用RT-PCR检测目的基因周期抑制蛋白P21和周期蛋白E2(CCNE2)mRNA的表达;Transwell体外侵袭实验检测细胞侵袭能力;蛋白印迹法检测蛋白表达;采用化学抑制剂或基因沉默法抑制胞内钙蛋白酶(CANP)活性。结果 E2(10 nmol/L)可刺激P21和CCNE2 mRNA表达显著增加(P<0.01);CANP抑制剂calpeptin(10μmol/L)或MEK抑制剂PD98059(10μmol/L)对E2诱导的P21和CCNE2 mRNA上调均有显著抑制作用(P<0.01);E2刺激细胞侵袭能力明显增强(P<0.01),而基因沉默CANP2明显抑制E2对CANP的激活及E2诱导的细胞侵袭效应(P<0.01)。结论 E2可通过激活胞内CANP诱导乳腺癌细胞P21和CCNE2 mRNA表达上调并促进细胞侵袭,提示CANP可能为抑制乳腺癌转移的一个潜在药物靶点。Objective To investigate the effect of 17β-estradiol（ E2） on the gene expression and coincident enhanced invasion,and the role of ERK-calpain signaling in the E2 action with the purpose of studying the molecular mechanism for the E2 effect. Methods MCF-7 breast cancer cells were used as a model system,RT-PCR was employed to evaluate the level of mRNA of target genes. Transwell invasion assay was applied to access cell invasive ability.Western blot was used to analyze the proteolysis of target proteins and siRNA was used to knockdown calpain2 gene expression and reduce the activity of calpain. Results Treatment of MCF-7 cells with E2 induced significant up-regulation of both P21 and CCNE2（ P〈 0. 01）,as well as enhanced invasion. Calpain or MEK specific inhibitors,calpeptin（ 10 μmol/L） and PD98059（ 10 μmol/L）,showed tremendous suppression of E2-stimulated expression of P21/CCNE2 mRNA（ P〈 0. 01）. Knockdown of calpain2 blocked E2-stimulated calpain activation and cell invasion（ P〈 0. 01）. Conclusions E2 induces up-regulation of P21/CNNE2 mRNA and enhances invasion and this effect is mediated via calpain,indicating calpain could be a potential target for the prevention of metastatic breast cancer.

关 键 词：雌激素 钙蛋白酶 侵袭 P21 周期蛋白E2 

分 类 号：R737.9[医药卫生—肿瘤]