基础研究(Basic Research)

miR-150-5p抑制肝癌细胞的迁移和侵袭及其机制

Published at: 2016年第25卷第8期

梁治坤 1 , 程凡天 1 , 胡走肖 1 , 郑小林 1
1 华中科技大学同济医学院附属武汉市中心医院 肝胆胰外科,湖北 武汉 430014
通讯作者 小林 郑 Email: xiaolingzheng999@sina.com
DOI: 10.3978/j.issn.1005-6947.10.3978/j.issn.1005-6947.2016.08.015
基金:

摘要

目的:探讨miR-150-5p在肝细胞癌(HCC)细胞迁移与侵袭中的作用及其调控机制。方法:用荧光定量PCR测定miR-150-5p在正常肝细胞系L02及HCC细胞系HepG2中的表达;将HepG2细胞分成两组,分别转染miR-150-5p(miR-150-5p组)与随机序列(对照组),转染后,分别用细胞划痕实验、Transwell小室基质渗透实验检测细胞的迁移和侵袭能力,用Western blot检测细胞基质金属蛋白酶2(MMP2)和基质金属蛋白酶9(MMP9)的蛋白表达。结果:miR-150-5p的表达量在HepG2细胞系中明显降低,为L02细胞系的0.26倍(P<0.01)。转染后,miR-150-5p组的miR-150-5p水平明显升高,为对照组的9.53倍(P<0.001);miR-150-5p组的细胞划痕愈合率明显低于对照组(54.63% vs. 87.51%,P<0.01),细胞侵袭数明显少于对照组(138个vs. 452个,P<0.01);MMP2与MMP9蛋白表达量均明显低于对照组(0.78 vs. 1.75;0.82 vs. 1.85,均P<0.05)。结论:miR-150-5p在HCC细胞中表达降低,升高miR-150-5p的表达可抑制HCC细胞的迁移和侵袭,机制可能与其下调MMP2和MMP9表达有关。


Inhibitory effect of miR-150-5p on migration and invasion of hepatocellular carcinoma cells and its mechanism

Abstract

Objective: To investigate the effect of miR-150-5p on migration and invasion in hepatocellular carcinoma (HCC) cells and the mechanism. Methods: The miR-150-5p expression in normal hepatic cells and HCC HepG2 cells was determined by real-time PCR. Then, the HepG2 cells were divided into two groups to transfect with miR-150-5p (miR-150-5p group) or scramble sequences (control group), respectively. After transfection, the cell migration and invasion abilities were measured by wound healing assay and Transwell invasion assay, and the protein expressions of matrix metalloproteinase 2 (MMP2) and 9 (MMP9) were examined by Western blot, respectively. Results: The expression level of miR-150-5p in HepG2 cells was significantly reduced, and was 26% of that in normal hepatic cells (P<0.01). After transfection, the miR-150-5p expression level in miR-150-5p group was significantly increased, which was 9.53 times of that in control group (P<0.001). In miR-150-5p group compared with control group, the healing rate of the scratch wound (54.63% vs. 87.51%, P<0.01) and number of invaded cells (138 vs. 452, P<0.01) were all significantly decreased. The protein expression levels of MMP2 and MMP9 (0.78 vs. 1.75; 0.82 vs. 1.85) were all significantly decreased (both P<0.05). Conclusion: The miR-150-5p expression is decreased in HCC cells, and up-regulation of miR-150-5p expression can inhibit the migration and invasion of HCC cells and the mechanism may be associated with its suppressing MMP2 and MMP9 expressions.


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引用

引用本文: 治坤 梁, 凡天 程, 走肖 胡, 小林 郑. miR-150-5p抑制肝癌细胞的迁移和侵袭及其机制[J]. 中国普通外科杂志, 2016, 25(8): 1175-1179.
Cite this article as: LIANG Zhikun, CHENG Fantian, HU Zouxiao, ZHENG Xiaolin . Inhibitory effect of miR-150-5p on migration and invasion of hepatocellular carcinoma cells and its mechanism[J]. Chin J Gen Surg, 2016, 25(8): 1175-1179.