长链非编码RNA TINCR通过miR-7调控肝癌细胞系Huh7侵袭和迁移  

Regulation of invasion and migration of cell line Huh7 by lncRNA TINCR via miR-7

作　　者：田小星 秦溧嫔 李茜[1] TIAN Xiao-xing;QIN Li-pin;LI Qian(Department of Infectious Diseases, the Fifth People's Hospital of Chongqing, Chongqing 400062, China)

机构地区：[1]重庆市第五人民医院感染性疾病科,重庆400062

出　　处：《基础医学与临床》2020年第8期1041-1046,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81804074)。

摘　　要：目的研究长链非编码RNA(lncRNA)组织分化诱导非蛋白质编码RNA(TINCR)通过miR-7调控肝癌Huh7细胞侵袭和迁移的机制。方法Real-time PCR测定Huh7细胞中TINCR表达。Huh7细胞中转染TINCR shRNA,CCK-8法测定增殖;Transwell小室法测定侵袭和迁移;Western blot测定MMP-2和MMP-9蛋白表达。生物信息学软件预测发现TINCR和miR-7有结合位点,荧光素酶报告系统鉴定二者靶向关系。Huh7细胞中共转染miR-7 inhibitor和TINCR shRNA,利用上述方法测定细胞增殖、迁移和侵袭变化。结果肝癌细胞中TINCR表达水平明显高于正常肝细胞(P<0.05)。转染TINCR shRNA后的Huh7细胞中TINCR水平降低(P<0.05),细胞增殖、迁移以及侵袭能力均下降(P<0.05),细胞中MMP-2和MMP-9蛋白水平也降低(P<0.05)。TINCR靶向负调控miR-7表达。miR-7 inhibitor可以提高下调TINCR后的肝癌细胞增殖、迁移、侵袭能力以及MMP-2、MMP-9蛋白表达水平(P<0.05)。结论下调TINCR可以通过靶向促进miR-7表达抑制肝癌细胞Huh7侵袭和迁移。Objective To study the mechanism of long non-coding RNA(lncRNA)TINCR to regulate the invasion and migration of Huh7 cells through miR-7.Methods The expression of TINCR was detected by real time PCR.TINCR shRNA was transfected and proliferation of Huh7 cells was determined by CCK-8;Invasion and migration were measured by Transwell chamber;The expression of MMP-2 and MMP-9 was detected by Western blot.Bioinformatics software predicted target relationship between TINCR and miR-7 and confirmed by luciferase report system.In Huh7 cells,miR-7 inhibitor and TINCR shRNA were co-transfected,the cell proliferation,migration and invasion were measured by the above methods.Results The expression level of TINCR in hepatoma cells was significantly higher than that in normal hepatocytes(P<0.05).After transfection of TINCR shRNA,the level of TINCR decreased in Huh7 cells(P<0.05),cell proliferation,migration and invasion all significantly decreased(P<0.05),MMP-2 and MMP-9 protein levels were also decreased(P<0.05).TINCR targeted negative regulation of miR-7 expression.miR-7 inhibitor can improve the proliferation,migration,invasion and the expression of MMP-2 and MMP-9 protein of hepatoma cells after down regulating TINCR(P<0.05).Conclusions Down regulation of TINCR may inhibit the invasion and migration of Huh7 cells through targeting at miR-7 expression.

关 键 词：肝癌 miR-7 迁移 组织分化诱导非蛋白质编码RNA 

分 类 号：R735.7[医药卫生—肿瘤]