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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。
Abstract：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.
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基本信息:
DOI：10.13417/j.gab.037.002708
中图分类号:R737.9
引用信息:
[1]席磊,付立新,唐曦,等.癌相关成纤维细胞miR-200c对乳腺癌细胞侵袭的影响[J].基因组学与应用生物学,2018,37(06):2708-2713.DOI:10.13417/j.gab.037.002708.
基金信息:
国家自然科学基金(81402180)资助
2018-06-25
2018-06-25