MicroRNA-485-5p抑制人结直肠癌细胞增殖和侵袭作用的研究-东南大学学报医学版

MicroRNA-485-5p抑制人结直肠癌细胞增殖和侵袭作用的研究

单位：1. 东南大学医学院, 江苏南京 210009;
2. 南京医科大学附属南京医院(南京市第一医院) 中心实验室, 江苏南京 210012

分类号：R735.3

出版年·卷·期（页码）：2018·37·第五期（814-819）

摘要：

目的：研究miR-485-5p在结直肠癌细胞中的表达水平，并探索其对结直肠癌细胞增殖和侵袭能力的影响。方法：运用荧光定量PCR法检测miR-485-5p在结直肠癌细胞株（HCT116和HCT8）及正常人类肠黏膜上皮细胞（FHC）中的表达水平。通过转染miR-485-5p mimic建立miR-485-5p过表达结直肠癌细胞，CCK-8实验及Transwell侵袭实验分别检测miR-485-5p对细胞增殖和侵袭能力的影响。生物信息学方法预测miR-485-5p下游靶基因并通过双荧光素酶报告基因检验、实时荧光定量PCR和Western blotting等方法进行实验验证。结果：miR-485-5p在结直肠癌细胞中的表达水平显著低于FHC细胞，差异有统计学意义（P<0.05）；miR-485-5p可抑制结直肠癌细胞在体外的增殖和侵袭能力；生物信息学结合双荧光素酶报告基因检测、实时荧光定量PCR及Western blotting等方法证实CD147为miR-485-5p调控的靶基因。结论：在结直肠癌中，miR-485-5p发挥了抑癌基因的功能，具有作为基因治疗的潜在价值。

Objective:To investigate the expression level of miR-485-5p in colorectal cancer cells and to explore its effect on the proliferation and invasion of colorectal cancer cells. Methods:The expression levels of miR-485-5p in human colorectal cancer cell lines (HCT116 and HCT8) and normal colon epithelial cell line (FHC) were detected by real-time fluorescence quantitative PCR. Cell proliferation and invasion ability was detected by Cell Counting Kit-8 assay (CCK-8) and Transwell assays. The downstream target gene of miR-485-5p was verified by dual-luciferase reporter assay, qRT-PCR and Western blotting. Results:The expression of miR-485-5p in human colorectal cancer cells was significantly lower than that in FHC cells (P<0.05). Experimental up-regulation of miR485-5p in HCT8 and HCT116 cells with its mimic could inhibit the proliferation and invasion of these cancer cells. The results of qRT-PCR and Western blotting showed that miR-485-5p inhibited the expression of CD147 by miR-485-5p, which was confirmed by bioinformatics and dual luciferase assay. Conclusion:In colorectal cancer,miR-485-5p plays a role of tumor suppressor gene by inhibiting the proliferation and invasion of colorectal cancer cells through CD147, it has the potential value as a gene therapy.

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