人乳腺癌微环境中癌相关成纤维细胞的生物学特征  

Biological characteristics of carcinoma-associated ibroblasts in human breast cancer microenvironment

作　　者：彭琼乐[1] 罗浩军[2] 孙艳[1] 韩明利[3] 王黎阳[1] 赵浏阳[1] 柳满然[1] 

机构地区：[1]重庆医科大学临床检验诊断学教育部重点实验室,重庆400016 [2]重庆医科大学附属第二医院三腺外科,重庆400016 [3]重庆医科大学附属第一医院内分泌乳腺外科,重庆400016

出　　处：《肿瘤》2013年第10期855-861,共7页Tumor

基　　金：国家自然科学基金资助项目(编号:81072147);教育部高校博士点基金资助项目(编号:20105503110001);教育部留学回国启动基金资助项目(编号:教外司[2011]508号);重庆医科大学检验医学院优秀博士基金(编号:070025000204)

摘　　要：目的:研究人乳腺癌微环境中癌相关成纤维细胞(carcinoma-associated ibroblasts,CAFs)的增殖、黏附、迁移、侵袭和凝胶收缩能力。方法:首先通过蛋白质印迹法检测细胞中纤黏蛋白(ibronectin,FN)、α-平滑肌肌动蛋白(alpha-smooth-muscle actin,α-SMA)和成纤维细胞活化蛋白(ibroblast activation protein,FAP)的表达来鉴别验证原代分离培养的CAFs与正常成纤维细胞(normal ibroblasts,NFs)。然后采用罗氏xCellingence活细胞分析系统、细胞计数法和CCK-8(cell counting kit-8)法比较CAFs与NFs的增殖差异。再通过细胞黏附实验、罗氏xCellingence分析系统、Transwell小室侵袭实验和凝胶收缩实验分别比较CAFs与NFs细胞的黏附、迁移、侵袭和收缩的能力。结果:原代分离培养的人乳腺癌CAFs细胞和NFs细胞均生长状态良好,增殖活跃,生长曲线的线型及趋势均符合细胞生长特点,但CAFs细胞的增殖能力明显高于NFs细胞。与NFs细胞相比,CAFs细胞的黏附、迁移、侵袭和收缩能力均明显增强。结论:人乳腺癌微环境中的CAFs细胞与NFs细胞的增殖与迁移存在明显差异,与NFs细胞相比,CAFs细胞具有较强的增殖、黏附、迁移、侵袭和收缩能力。Objective： To investigate the proliferation, adhesion, migration, invasion, and contraction capacities of carcinoma-associated fibroblasts （CAFs） in human breast cancer microenvironment. Methods： The protein expressions of fibronectin （FN）, alpha-smooth-muscle actin （^-SMA） and fibroblast activation protein （FAP） were detected by Western blotting, so as to distinguish CAFs cells from normal fibroblasts （NFs）. The proliferation of CAFs and NFs was detected by Roche xCellingence system, cell counting, and cell counting kit-8 （CCK-8） assay. The adhesion, migration, invasion and contraction capacities of CAFs were evaluated by the cell adhesion experiment, Roche xCellingence system, Transwell invasion assay and collagen gel contraction assay, respectively. Results： The primary CAFs and NFs cells which were isolated from human breast cancer grew in good condition with active proliferation. The linear types and trends of their growth curves were accorded with the cell growth characteristics. While compared with NFs, CAFs had a robust proliferation capacity, and the obviously stronger abilities of adhesion, migration, invasion, and contraction. Conclusion： There are prodigious differences of proliferation and migration between CAFs and NFs cells in human breast cancer microenvironment. CAFs have the stronger abilities of proliferation, adhesion, migration, invasion, and contraction than NFs.

关 键 词：乳腺癌 成纤维细胞 肿瘤相关 肿瘤形成过程 细胞增殖 细胞黏附 肿瘤侵润 细胞收缩 

分 类 号：R730.231[医药卫生—肿瘤]