鼻咽癌相关成纤维细胞miRNA表达差异

Expression of miRNA in Nasopharyngeal Carcinoma-Associated Fibroblasts

[目的]研究人鼻咽癌微环境癌相关成纤维细胞(CAF)与正常成纤维细胞(NF)miRNA表达的差异。[方法]运用1型胶原酶消化法处理正常鼻咽黏膜组织标本,原代分离培养NF细胞,与TGF-β1共孵育后获得鼻咽癌CAF细胞;利用Transwell实验比较CAF与NF细胞的侵袭能力;利用miRNA芯片分析CAF与NF细胞miRNA表达的差异。[结果]成功分离得到原代培养的CAF与NF细胞,与NF细胞相比,CAF细胞的侵袭能力明显增强;miRNA芯片分析结果显示,CAF有13个异常表达miRNA(P<0.05),其中miR-221-5p、miR-155、miR-210上调表达,10个下调表达(miR-205、miR-200b、miR-342-3p、miR-329、miR-487a、miR-455-3p、miR-301a、miR-652、miR-432、let-7g)。[结论 ]人鼻咽癌微环境CAF的miRNA表达谱发生显著变化。

[Purpose] To investigate the differential expression of miRNAs between nasopharyngeal carcinoma-associated fibroblasts（CAF） and normal fibroblasts（NF）. [Methods] The primary NF cells were isolated from nasopharyngeal epithelial tissue by collagenase digestion method,and CAF cells were obtained from nasopharyngeal carcinoma specimen after incubation with TGF β-1. Cell invasion ability was examined by transwell assay,and the expression of miRNAs was detected by microarray analysis. [Results] The primary cultures of CAF and NF cells were successfully isolated.Compared with NF cells,the invasion ability of CAF cells was markedly increased. Micro RNA microarray analysis showed that 13 miRNAs were expressed differentially between NF and CAF cells（P〈0.05）,of which 3 were up-regulated（miR-221-5p,miR-155,mir-210） and 10 were down-regulated（mir-205,mir-200 b,mir-342-3p,miR-329,mir-487 a,miR-455-3p,which,miR-652,miR-432,let-7g）.[Conclusion] The expression profile of miRNAs in human nasopharyngeal carcinoma-associated fibroblasts is significantly changed.