lncRNA SNHG5在缺氧诱导肝细胞癌细胞侵袭及迁移中的作用

Role of lncRNA SNHG5 in the migration and invasion of hypoxia-induced hyhepa-tocellular carcinoma cells

目的:探讨长链非编码RNA(long non-coding RNA,lncRNA)SNHG5在缺氧诱导肝细胞癌(hepatocellular carcinoma,HCC)细胞侵袭及迁移中的作用。方法:收集2017年1月至2018年6月西安交通大学第一附属医院手术切除的20例HCC患者的癌与癌旁组织标本,以及人HCC细胞系HepG2、MHCC-97L、MHCC-97H、Huh7和永生化人肝细胞LO2。利用生物信息学方法分析缺氧诱导因子1α(hypoxia-inducible factor 1α,HIF-1α)与SNHG5的结合位点,将pCMV-HIF-1α、shRNA-SNHG5(sh-SNHG5)质粒转染进HCC细胞,用qPCR法检测HCC组织和缺氧条件下HCC细胞中SNHG5的表达水平,用Western botting检测HCC细胞中HIF-1α蛋白的表达水平,用Transwell小室法检测沉默SNHG5后常氧和缺氧条件下对HCC细胞侵袭及迁移能力的影响。结果:分别与癌旁组织和LO2细胞比较,HCC组织和各细胞系中SNHG5表达水平显著上调(均P<0.01)。缺氧可上调HCC细胞中SNHG5的表达水平,其机制可能与缺氧活化HIF-1α与SNHG5启动子结合从而促进其转录有关;缺氧能够增强HepG2和MHCC-97L细胞的侵袭及迁移能力(均P<0.01),沉默SNHG5表达能够显著抑制缺氧条件下HepG2和MHCC-97L细胞的侵袭及迁移能力(均P<0.01)。结论:SNHG5在HCC组织及细胞系中高表达,并在缺氧诱导的HCC细胞侵袭及迁移中发挥重要作用。

Objective:To explore the regulatory effect of long non-coding RNA(lncRNA)SNHG5 on invasion and migration of hypoxia-induced hepatocellular carcinoma(HCC)cells.Methods:A total of 20 pairs of cancer and para-cancerous tissue specimens resected from HCC patients in the First Affiliated Hospital of Xi'an Jiaotong University from January 2017 to June 2018,and human HCC cell lines(HepG2,MHCC-97L,MHCC-97H,Huh7)as well as immortalized human liver LO2 cells were collected for this study.Bioinformatics methods were used to analyze the binding sites between hypoxia-inducible factor 1α(HIF-1α)and SNHG5.pCMVHIF-1αand shRNA-SNHG5(sh-SNHG5)plasmids were transfected into HCC cells,respectively.qPCR was used to detect the expres-sion level of SNHG5 in HCC tissues and hypoxia-induced HCC cells.Western botting was used to detect the expression level of HIF-1αprotein in HCC cells,and Transwell chamber method was used to detect the migration and invasion ability of HCC cells after SNHG5 si-lence under normoxia and hypoxia condition.Results:Compared with para-cancerous tissues and immortalized human liver LO2 cells,the expression of SNHG5 was significantly up-regulated in HCC tissues and cell lines(all P<0.01).Hypoxia promoted the expression level of SNHG5 in HCC cells,and its mechanism might be related to the combination of hypoxia-activated HIF-1αand SNHG5 promoter to promote its transcription.Hypoxia promoted the invasion and migration ability of HepG2 and MHCC-97L cells(all P<0.01),but knockdown of SNHG5 significantly inhibited the invasion and migration ability of HepG2 and MHCC-97L cells under hy-poxic conditions(all P<0.01).Conclusion:SNHG5 is highly expressed in HCC tissues and cell lines and plays an important role in the invasion and migration of HCC cells induced by hypoxia.