乳腺癌中肿瘤相关成纤维细胞原代分离、培养及相关生物学特性分析  

Primary isolation, culture and biological property analysis of breast cancer-associated fibroblast

作　　者：张春敏[1] 郭变琴[1] 吴立翔[1] Zhang Chunmin Guo Bianqin Wu Lixiang(Department of Clinical Laboratory, Chongqing Cancer Institute, Chongqing Cancer Hospital, Chongqing Cancer Center, Chongqing 400030, Chin)

机构地区：[1]重庆市肿瘤研究所重庆市肿瘤医院重庆市癌症中心检验科,400030

出　　处：《肿瘤研究与临床》2017年第8期510-514,共5页Cancer Research and Clinic

基　　金：重庆市科学技术委员会科技计划（cstc2014jcyA1633）

摘　　要：目的 探索乳腺癌中肿瘤相关成纤维细胞（CAF）稳定可行的原代分离、培养方法,探讨CAF的生物学特性.方法 取10例未进行任何放疗、化疗及内分泌治疗的手术切除乳腺癌组织及其癌灶外缘2 cm处的配对癌旁组织,采用组织块酶消化法分离培养CAF.流式细胞术检测细胞周期,细胞免疫化学法检测CAF生物学标志波形蛋白（Vimentin）、α平滑肌肌动蛋白（α-SMA）以及细胞增殖指标Ki-67表达情况,Transwell小室侵袭、迁移实验检测CAF侵袭、迁移能力.结果 通过条件优化,成功分离培养CAF.细胞免疫化学结果提示,CAF高表达Vimentin、α-SMA以及Ki-67.流式细胞术结果显示,CAF较乳腺癌旁成纤维细胞S期细胞比例增加,提示CAF增殖能力强,分裂增殖活跃.CAF具有很强的侵袭、迁移能力[CAF组和乳腺癌旁成纤维细胞组侵袭细胞数分别为（79.7±3.5）、（66.3±1.5）个,迁移细胞数分别为（242.3±3.1）、（218.3±2.1）个,均P〈0.05].结论 优化组织块酶消化法并成功分离、培养CAF.CAF具有较强的分裂增殖能力和侵袭迁移能力.Objective To explore a stable and feasible method for the primary culture of breast cancer-associated fibroblast （CAF）, and to offer the theoretical basis for the further studies about the role of CAF in the processing of breast cancer by analyzing the biological property of CAF. Methods CAF was primary cultured with tissue block enzymolytic method. Flow cytometry （FCM） was used to detect the cell cycle. The expression of Vimentin, α-SMA and Ki-67 were detected by cell immunochemistry method. Transwell assay was used to detect the invasion and migration capacity of CAF. Results The CAF was successfullyculturedbyconditionoptimization.ImmunohistochemicalresultsshowedthatVimentin, α-SMA and Ki-67 were highly expressed in CAF.Comparedwithfibroblastsfrombreastcancersidestissues,the proliferating ability and the invasion and migration capacities of CAF were enhanced （number of invasive cells： 79.7 ± 3.5 vs. 66.3 ± 1.5, number of migrated cells： 242.3 ± 3.1 vs. 218.3 ± 2.1, both P〈0.05）. Conclusion CAF cultured from breast cancer has better proliferating ability and strong invasion and migration capacities.

关 键 词：乳腺肿瘤 成纤维细胞 细胞培养技术 肿瘤侵润 细胞运动 

分 类 号：R737.9[医药卫生—肿瘤]