雌二醇通过钙离子/钙激活中性蛋白酶通路诱导乳腺癌细胞迁移及FAK蛋白剪切  

Estradiol Induces Cell Migration and Proteolysis of Focal Adhesion Kinase (FAK) via Calcium /Calpain Signaling in Breast Cancer Cells

作　　者：武春兰[1,2] 李阳[1] 刘晓红[1] 侯建美[1] 朱筑霞[1] 杨莉[1] 王旭东[1] 

机构地区：[1]贵阳医学院生理学教研室,贵阳550004 [2]贵阳中医学院第一附属医院科研实验室

出　　处：《肿瘤防治研究》2012年第9期1041-1045,共5页Cancer Research on Prevention and Treatment

基　　金：国家自然科学基金资助项目(30860093);贵州省优秀科技教育人才省长专项基金资助项目[黔省专合字(2008)56号]

摘　　要：目的观察雌二醇(E2)诱导MCF-7乳腺癌细胞迁移和局部黏着斑激酶(FAK)蛋白剪切及钙离子(Ca2+)/钙激活中性蛋白酶(CANP)通路在E2效应中的介导作用,探讨E2诱导细胞迁移的信号机制。方法以人乳腺肿瘤细胞系MCF-7为体外研究模型;采用蛋白印迹法分析细胞FAK的蛋白剪切效应;通过伤口愈合实验观察细胞迁移;采用CANP抑制剂(Calpeptin)或胞内钙螯合剂(BAPTA)预处理细胞,观察其对E2诱导细胞迁移及FAK蛋白剪切的影响。结果 E2可明显诱导MCF-7细胞迁移同时FAK出现蛋白剪切,此效应可被Calpeptin或BAPTA预处理显著抑制;E2还可诱导CANP1自身蛋白剪切,也可被Calpeptin或BAPTA显著削弱或阻断。结论 E2可通过细胞内Ca2+/CANP信号通路诱导乳腺癌细胞迁移及FAK蛋白剪切,后者可能在E2诱导的细胞迁移效应中起重要作用。Objective To investigate the role and signaling mechanisms of calcium-activated neutral protease（CANP）involved in cell migration and proteolysis of FAK protein induced by estradiol（E2）in breast cancer.Methods Wound healing assay was carried out to evaluate cell migration in human breast cancer cell line MCF-7.The effects of E2or epidermal growth factor（EGF）on the truncation of focal adhesion kinase（FAK）protein were detected by Western blotting.A specific inhibitor of CANP,Calpeptin,and an intracellular calcium chelator,BAPTA/AM,were applied to pre-treat cultured cells to evaluate their influences on the E2-stimulated cell migration and FAK proteolysis.Results E2was able to markedly induced MCF-7cell migration and FAK proteolysis.This effect can be significantly inhibited by Calpeptin BAPTA pre-treatment.E2also induced CANP1autolysis,which might be significantly weakened or blocked by Calpeptin or BAPTA.Conclusion E2induced breast cancer cell migration and FAK proteolysis（an important factor in E2-induced cell migration）through intracellular Ca2＋/the CANP signal pathway.

关 键 词：雌激素 乳腺癌 钙离子 卡配因 细胞迁移 局部黏着斑激酶 

分 类 号：R737.9[医药卫生—肿瘤] R730.231[医药卫生—临床医学]