FAK在雌激素诱导MCF-10A乳腺上皮细胞转化中的表达及其意义  

Altered expression of FAK in estrogen-transformed MCF-10A mammary epithelial cells and its significance*

作　　者：杨莉[1] 李阳[1] 朱筑霞[1] 王旭东[1] 

机构地区：[1]贵阳医学院生理学教研室,550004

出　　处：《重庆医学》2013年第20期2376-2377,2408,共3页Chongqing medicine

基　　金：国家自然科学基金资助项目(30860093);贵州省科技教育优秀人才专项基金资助项目[黔省专合字2008-56]

摘　　要：目的观察17-β雌二醇(E2)诱导雌激素受体(ER)阴性乳腺上皮细胞MCF-10A转化及伴随的局部黏着斑激酶(FAK)表达变化,探讨FAK在细胞转化中的生物标记意义。方法以ER阴性MCF-10A细胞为研究模型,E2连续刺激模型细胞5周(共传11代)建立转化细胞模型,采用蛋白印迹法检测蛋白表达变化、软琼脂集落形成实验检测非贴壁依赖集落形成、伤口愈合实验检测细胞迁移及细胞胎盼蓝染色计数观察细胞生长。结果 E2连续刺激MCF-10A细胞5周后,细胞生长速度明显增快、接触抑制性消失,细胞迁移显著增快,同时在软琼脂基质中形成集落。在E2诱导细胞转化早期即出现FAK蛋白表达增强及蛋白剪切,但乳腺癌标记分子周期蛋白E表达无明显变化。结论 E2诱导ER阴性MCF-10A乳腺上皮细胞转化中始终伴随FAK表达变化,提示FAK可能成为预测ER阴性乳腺细胞转化的早期生物标记物。Objective To investigate 17β-estradiol（E2）-induced transformation and coincident altered expression of focal adhe- sion kinase（FAK） in estrogen receptor（ER）-negative mammary epithelial cell line MCF-10A,so as to understand whether FAK is a potential biomarker in E2-transformed mammary cells. Methods MCF-10A cells were treated with E2 for 5 weeks（ll passages） to establish an E2-transformed cell model. Western blotting was used to access protein expression, soft-agar colony formation test was employed to observe anchorage- independent growth, wound healing assay was applied to observe cell migration, and Trypan blue staining and cell counting was employed to observe cell growth. Results After treatment of wild type MCF-10A cells with E2 for 5 weeks, the cells displayed increased proliferation, migration, and colony formation, indicating a transformed phenotype. More impor- tantly,enhanced expression and proteolysis of FAK was noticed during the progression of E2-induced transformation. Conclusion E2-induced transformation of ER-negative MCF-10A breast epithelial cells is accompanied by altered expression of FAK, indicating FAK as a potential early biomarker for transformed ER-negative breast cells.

关 键 词：17—β雌二醇 黏着斑激酶 乳腺上皮细胞 细胞转化 生物标记物 

分 类 号：R737.9[医药卫生—肿瘤]