钙蛋白酶在乳腺上皮转化细胞对雌激素刺激反应中的作用  

The role of calpain in the responsiveness of transformed mammary epithelial cells to estrogen stimulation

作　　者：杨莉[1] 朱筑霞[1] 刘晓红[1] 胡晓霞[1] 王红梅[1] 王旭东[1] 

机构地区：[1]贵阳医学院生理学教研室,贵州贵阳550004

出　　处：《基础医学与临床》2013年第7期819-823,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(30860093);贵州省科技教育优秀人才省长专项基金(黔省专合字2008-56)

摘　　要：目的探讨经雌激素(E2)转化的MCF-10A乳腺上皮细胞对E2刺激的敏感性及细胞内钙蛋白酶(CANP)在E2效应中的介导作用,以深入了解E2的信号传导机制。方法以E2转化的MCF-10A乳腺上皮细胞为研究模型、用E2(50 nmol/L)刺激细胞,以蛋白印迹法观察蛋白分子质量变化,以CANP特异性底物局部黏着激酶(FAK)蛋白剪切作为CANP激活的指标,伤口愈合实验观察细胞迁移变化,CANP抑制剂预处理观察CANP在E2效应中的作用。结果转化细胞CANP活性明显高于非转化细胞,表现为前者出现FAK明显蛋白剪切,而后者FAK主要为野生型(125 ku),同时转化细胞迁移明显增强(P<0.01)。E2(10 nmol/L)刺激转化细胞可进一步促进FAK蛋白剪切和细胞迁移(P<0.01),而非转化细胞对E2刺激不敏感。CANP抑制剂-1(ALLN)可有效阻断E2刺激转化细胞FAK蛋白剪切及细胞迁移(P<0.01)。结论转化细胞对E2刺激的敏感性增加,其机制可能与胞内CANP活性增强有关,提示在转化细胞存在活跃的E2-CANP-FAK信号活动。Objective To inves cells to E2 stimulation and the tigate the sensitirity of 17 [3estradiol （E2）-transformed MCF-10A mammary epithelial role of calpain （CANP） in the E2-induced effect. Methods E2-transformed MCF- 10A mammary epithelial cells were prepared and used as a model system. Western blotting was employed to observe proteolysis of focal adhesion kinase （FAK） as a marker of calpain activity, wound healing assay was performed to investigate cell migration. Calpain inhibitor was used to find role of calpain in the E2-induced effect. Results In non-transformed MCF-10A cells, focal adhesion kinase （FAK） was primarily expressed in the form of wild type, while in E2（50 nmol/L）-transformed cells FAK was significantly proteolyzed, indicating increased activity of cal- pain. Furthermore, transformed cells showed increased migration as compared with control （P 〈0.01 ）. Treatment of transformed cells with E2 （ 10 nmol/L） triggered further truncation of FAK and enhancement of migration （P 〈0. 01 ）, while non-transformed cells were resistant to E2 stimulation. Additionally, pre-treatment of trans- formed cells with calpain inhibitor-1 （ALLN, 10 μmol/L） abrogated E2-enhanced FAK processing and migration（P 〈0.01 ）. Conclusions Transformed MCF10-A mammary epithelial cells displays increased responsiveness to E2 stimulation, and this effect may be mediated through activation of calpain, indicating an active E2-CANP-FAK signaling in the transformed cells.

关 键 词：17Β-雌二醇 细胞转化 敏感性 卡配因 乳腺上皮细胞 

分 类 号：R737.9[医药卫生—肿瘤]