基础医学与临床 ›› 2020, Vol. 40 ›› Issue (8): 1041-1046.

• 研究论文 • 上一篇    下一篇

长链非编码RNA TINCR通过miR-7调控肝癌细胞系Huh7侵袭和迁移

田小星, 秦溧嫔, 李茜*   

  1. 重庆市第五人民医院 感染性疾病科, 重庆 400062
  • 收稿日期:2019-12-19 修回日期:2020-03-29 出版日期:2020-08-05 发布日期:2020-07-29
  • 通讯作者: *lwdvd6@163.com
  • 基金资助:
    国家自然科学基金(81804074)

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

TIAN Xiao-xing, QIN Li-pin, LI Qian*   

  1. Department of Infectious Diseases, the Fifth People's Hospital of Chongqing, Chongqing 400062, China
  • Received:2019-12-19 Revised:2020-03-29 Online:2020-08-05 Published:2020-07-29
  • Contact: *lwdvd6@163.com

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。

关键词: 肝癌, miR-7, 迁移, 组织分化诱导非蛋白质编码RNA

Abstract: 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.

Key words: hepatocellular carcinoma, miR-7, migration, tissue differentiation inducing non-protein coding RNA

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