癌相关成纤维细胞对人舌鳞状细胞癌上皮-间质转化的影响

摘要/Abstract

摘要： 目的:探讨癌相关成纤维细胞(CAFs)对舌鳞癌细胞侵袭、转移的作用及可能机制。方法:应用胶原酶消化法原代分离舌癌相关成纤维细胞及对应正常成纤维细胞(NFs),采用Western免疫印迹、ELISA、激光共聚焦和实时定量PCR分别检测vimentin、cytokeratin、α-SAM和SDF1蛋白质及mRNA水平表达差异。以舌鳞状细胞癌SCC9为研究对象,建立共培养体系,采用Western免疫印迹、激光共聚焦和实时定量PCR检测SCC9中上皮-间质转化标志蛋白E-cadherin、vimentin和fibronectin及其mRNA水平的变化,Transwell实验检测细胞侵袭、迁移能力。采用SPSS13.0软件包对数据进行统计学分析。结果:胶原酶消化法成功获取CAFs及对应NFs,CAFS和NFs均表达vimentin,不表达cytokeratin。其中,CAFs高表达α-SAM并分泌SDF1(P<0.01)。与SCC9共培养2周后,SCC9呈梭形改变,E-cadherin表达降低,vimentin和fibronectin升高(P<0.01),同时其迁移能力增强。结论:胶原酶消化法能成功获取原代CAFs。CAFs可通过诱导SCC9上皮-间质转化进而促进其侵袭、转移。

Abstract: PURPOSE: To examine the role of cancer-associated fibroblasts (CAFs) in promoting invasion of tongue squamous cell carcinoma (TSCC) and the possible mechanism. METHODS: A series of matched pairs of CAFs and normal fibroblasts (NFs) were primarily cultured in collagenase digestion. Identified markers of alpha smooth muscle actin (α-SMA), stromal cell-derived factor-1(SDF1), vimentin and cytokeratin (CK) were observed. Transwell assay, Western blot, reverse transcription-PCR and immunofluorescence staining were carried out to assess the role of CAFs in promoting migration, invasion and inducing epithelial to mesenchymal transition (EMT) of SCC9, compared with NFs. RESULTS: Both primarily cultured CAFs and NFs expressed vimentin, but not CK. CAFs showed significantly higher α-SMA expression and SDF1 secretion, as well as mRNA level of α-SMA and SDF1. Co-culture with CAFs induced up-regulation of EMT markers (fibronectin and vimentin) and down-regulation of E-cadherin of SCC9, as well as enhanced invasion. CONCLUSION: Collagenase digestion is a reliable way to primary culture of CAFs. CAFs may induce EMT markers and functional changes in SCC9. Supported by National Natural Science Foundation of China (81172563).

Key words: Cancer-associated fibroblast, Tumor microenvironment, Tongue squamous cell carcinoma, Epithelial-mesenchymal transition

中图分类号:

R739.8

周斌,陈伟良,王友元,林钊宇,张大明,范松. 癌相关成纤维细胞对人舌鳞状细胞癌上皮-间质转化的影响[J]. 中国口腔颌面外科杂志, 2014, 12(3): 198-204.

ZHOU Bin, CHEN Wei-liang, WANG You-yuan, LIN Zhao-yu, ZHANG Da-ming, FAN Song. Cancer-associated fibroblasts induce epithelial-to-mesenchymal transition in tongue squamous cell carcinoma[J]. China J Oral Maxillofac Surg, 2014, 12(3): 198-204.

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