微小RNA-646/CXC型趋化因子配体10轴对肝癌侵袭及其相关蛋白的影响  

Effect of microRNA-646/CXC tchemokine ligand 10 axis on invasion and related proteins of hepatocellular carcinoma

作　　者：王哲[1] 张朕 冯延冰 刘蕾[4] 裴正浩 乔兵兵[5] Wang Zhe;Zhang Zhen;Feng Yanbing;Liu Lei;Pei Zhenghao;Qiao Bingbing(Department of Liver Surgery,Nanyang Central Hospital Affiliated to Zhengzhou University,Nanyang 473000,China;Department of Liver General Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China;Department of Emergency Surgery,Nanyang Central Hospital,Zhengzhou University,Nanyang 473000,China;Department of Imaging,Nanyang Central Hospital,Zhengzhou University,Nanyang 473000,China;Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学附属南阳市中心医院肝脏外科,473000 [2]郑州大学第一附属医院普外科,450000 [3]郑州大学附属南阳市中心医院急诊外科,473000 [4]郑州大学附属南阳市中心医院影像科,473000 [5]郑州大学第一附属医院肝胆胰外科,450000

出　　处：《中华实验外科杂志》2020年第9期1720-1723,共4页Chinese Journal of Experimental Surgery

摘　　要：目的观察微小RNA(miRNA,miR)-646/CXC型趋化因子配体10(CXCL10)轴对肝癌侵袭的影响并探讨其分子机制。方法选取2018年6月到2020年1月南阳市中心医院收集的108例肝癌组织和对应癌旁组织作为研究对象,采用荧光定量聚合酶链反应(PCR)分析miR-646表达水平。采用Lipofectamine 2000转染miR-646 mimic和对照mimic至肝癌细胞MHCC97H,转染48 h后采用Transwell分析细胞侵袭能力;采用生物信息学和双荧光素酶报告基因分析miR-646的靶基因,采用蛋白质印迹法(Western blot)分析细胞和肿瘤组织中CXCL10蛋白表达水平。组间数据比较采用t检验。结果与癌旁组织(1.10±0.19)比较,肝癌组织中miR-646表达水平(0.44±0.17)显著下调,差异有统计学意义。与对照组细胞[(108.55±12.35)个]比较,实验组细胞侵袭数量[(45.36±8.99)个]显著下调,差异有统计学意义。生物信息学和双荧光素酶报告基因显示CXCL10是miR-646的靶基因。与癌旁组织(0.68±0.10)比较,肝癌组织中CXCL10表达水平(1.33±0.21)显著上调,差异有统计学意义。与对照组细胞(0.54±0.11)比较,实验组细胞表CXCL10表达水平(0.21±0.09)显著下调,差异有统计学意义。与癌旁组织基质金属蛋白酶(MMP)-2和MMP-9蛋白表达水平(0.85±0.19和0.58±0.18)比较,肝癌组织中MMP-2和MMP-9表达水平(1.75±0.25和1.49±0.18)显著增加,差异有统计学意义。与对照组细胞MMP-2和MMP-9表达水平(1.24±0.14、1.16±0.18)比较,实验组细胞MMP-9和MMP-2表达水平(0.37±0.12、0.18±0.08)显著下调,差异有统计学意义(t=3.018、5.091,P<0.05)。结论miR-646通过靶向CXCL10调节肝癌细胞的侵袭能力。Objective To investigate the effect of microRNA(miRNA,miR)-646/CXC tchemokine ligand 10(CXCL10)axis on invasion of hepatocellular carcinoma and its molecular mechanism.Methods 108 cases of liver cancer tissues and adjacent tissues from June 2018 to January 2020 were selected as the research objects.The expression level of miR-646 was analyzed by fluorescence quantitative polymerase chain reaction(PCR).miR-646 mimics and control mimics were transfected into hepatoma cell line MHCC97H by Lipofectamine 2000.After 48 hours of transfection,the invasion ability of the cells was analyzed by Transwell.The target gene of miR-646 were analyzed by bioinformatics and double luciferase reporter genes.The expression level of CXCL10 protein and invasion associated proteins in cells and tumor tissues were analyzed by Western blotting.T test was used to compare the data between groups,and the difference was statistically significant(P<0.05).Results Compared with the adjacent tissues(1.10±0.19),the expression level of miR-646 in hepatocellular carcinoma(HCC)tissues(0.44±0.17)significantly downregulated.Compared with the control group[(108.55±12.35)cells],the number of invasive cells in the experimental group[(45.36±8.99)cells]significantly decreased.Bioinformatics and double luciferase reporter gene showed that CXCL10 was the target gene of miR-646.Compared with adjacent tissues(0.68±0.10),the expression level of CXCL10 in HCC tissues(1.33±0.21)significantly up-regulated.Compared with the control group(0.54±0.11),the expression level of CXCL10 in the experimental group(0.21±0.09)significantly reduced.Compared with the expression levels of matrix metalloproteinase(MMP)-2 and MMP-9 protein in adjacent tissues(0.85±0.19 and 0.58±0.18),the expression levels of MMP-2 and MMP-9 in HCC tissues(1.75±0.25 and 1.49±0.18)significantly increased.Compared with the control group(1.24±0.14,1.16±0.18),the expression levels of MMP-9 and MMP-2 in the experimental group(0.37±0.12,0.18±0.08)significantly decreased.Conclusion MiR-646 r

关 键 词：微小RNA-646 肝癌 侵袭 CXC型趋化因子配体10 

分 类 号：R735.7[医药卫生—肿瘤]