摘要
探讨miR-200c在癌相关成纤维细胞（cancer-associated fibroblasts,CAF）活化中的作用,研究其对人乳腺癌细胞MDA-MB-231侵袭能力的影响。通过质粒构建的方法分别获得miR-200c稳定干扰的NF（normal fibroblast）及其对照细胞株、miR-200c稳定过表达的CAF及其对照细胞株;采用Western blotting法检测miR-200c对CAF活化标志物α-SMA和FAP表达的影响;采用细胞划痕愈合实验检测miR-200c对CAF迁移能力的影响;采用胶收缩实验和侵袭实验分别检测miR-200c对CAF细胞外基质和对癌细胞侵袭能力的影响。成功构建了miR-200c稳定干扰的NF及其对照细胞株、miR-200c稳定过表达的CAF及其对照细胞株;与低表达miR-200c的细胞株相比,高表达miR-200c细胞株的α-SMA和FAP表达水平及迁移能力明显降低,其胶收缩能力和癌细胞的侵袭能力也明显减弱。miR-200c能够明显抑制CAF细胞的活化并通过细胞外基质重塑（extracellular matrix remodeling,ECM）的方式抑制癌细胞的侵袭。
To investigate the effect of miR-200c in cancer-associated fibroblast（CAF） on the invasion of human breast cancer cell MDA-MB-231. Normal fibroblast（NF） lowly expressing miR-200c, CAF highly expressing miR-200c and the control cell lines were established by the method of plasmid construction. The expression of α-SMA and FAP, activation protein marker of CAFs, were detected by western blot and the effects of miR-200c on migration of CAFs were detected by cell wound healing assay. The influence of miR-200c on extracellular matrix remodeling（ECM） of CAF and on the invasion of breast cancer cells were tested by collagen-remodeling assay and cell invasion assay, respectively. NF lowly expressing miR-200c, CAF highly expressing miR-200c and the control cell lines were well established. The expression of α-SMA and FAP, migration of CAF, collagen shrinking ability and the invasion of breast cancer cell MDA-MB-231 in cell lines highly expressing miR-200c were remarkably lower than those in cell lines lowly expressing miR-200c. miR-200c of CAF inhibits the invasion ability of breast cancer cell MDA-MB-231 by means of inhibition of CAF activation and extracellular matrix remodeling.
作者
席磊
付立新
唐曦
杜艳娥
杨丹
陈燕林
柳满然
侯懿烜
Xi Lei;Fu Lixin;Tang Xi;Du Yan＇e;Yang Dan;Chen Yanlin;Liu Manran;Hou Yixuan(Key Laboratory of Medical Diagnostics of Ministry of Education, Department of Laboratory Medicine, Chongqing Medical University, Chongqing, 400016;The First Affiliated Hospital of Chongqing Medical University, Chongqing, 400016)
出处
《基因组学与应用生物学》
CAS
CSCD
北大核心
2018年第6期2708-2713,共6页
Genomics and Applied Biology
基金
国家自然科学基金(81402180)资助
关键词
乳腺肿瘤
癌相关成纤维细胞
细胞外基质重塑
肿瘤侵袭
Breast cancer
Cancer-associated fibroblast
Extracellular matrix remodeling
Cancer invasion
