MicroRNA-124靶向调控caveolin-1介导肝癌细胞恶性行为的研究

MicroRNA-124 mediates hepatocellular carcinoma malignancy by targeting caveolin-1

目的:检测miR-124、caveolin-1在肝癌组织及细胞系中的表达,探讨miR-124靶向调控caveolin-1对肝癌细胞增殖和侵袭的影响。方法:回顾性分析2012年8月至2014年7月32例于大连医科大学附属第一医院收治的行肝癌切除术患者的临床资料和标本,通过实时定量逆转录聚合酶链反应(qRT-PCR)检测肝癌、癌旁组织以及肝癌细胞系中miR-124、caveolin-1的表达;通过靶基因预测及双荧光素酶报告分析miR-124与caveolin-1的靶向关系;通过qRT-PCR、Western blot检测miR-124对caveolin-1表达的调控;分别用CCK8、平板克隆及Transwell检测细胞增殖和侵袭能力;通过绘制生存曲线,分析miR-124及caveolin-1表达与临床肝癌患者预后的相关性。结果:miR-124在肝癌组织中的表达低于癌旁组织,在高转移肝癌细胞MHCC97H中的表达低于低转移肝癌细胞MHCC97L,而caveolin-1呈现相反的趋势;caveolin-1为miR-124的靶基因,调控miR-124可影响caveolin-1水平;MHCC97H中导入miR-124 mimic可抑制该细胞增殖及侵袭能力,而上调caveolin-1促进该细胞增殖及侵袭能力;敲低低转移肝癌细胞MHCC97L中miR-124水平可增强该细胞的增殖及侵袭能力,下调caveolin-1抑制该细胞的增殖及侵袭能力;肝癌组织miR-124高水平、caveolin-1低水平的患者5年生存时间显著长于相应的miR-124低水平组、caveolin-1高水平组。结论:miR-124通过靶向调控caveolin-1介导肝癌的增殖与侵袭。

Objective:To explore the effect of microRNA-124(miR-124)/caveolin-1 in the progression of hepatocellular carcinoma(HCC).Methods:The expression levels of miR-124 and caveolin-1 were evaluated in HCC tissues,adjacent tissues,and cell lines by qRTPCR.A dual-luciferase reporter gene assay was used to confirm the relationship between microRNA-124 and caveolin-1.MicroRNA-124 mediated caveolin-1 levels,which was determined by qRT-PCR and Western blot analyses.CCK-8,colony formation,and transwell assays were used to evaluate the proliferation and invasion of HCC cells.Results:A low level of miR-124 together with a high level of caveolin-1 was found in HCC tissues and MHCC97 H cells compared with control cells.Caveolin-1 was determined to be the target gene of miR-124,and alterations of miR-124 could modulate caveolin-1.Cells transfected with a miR-124 mimic in MHCC97 H cells inhibited proliferation and invasion,whereas upregulation of caveolin-1 promoted proliferation and invasion.MHCC97 L cells transfected with a miR-124 inhibitor showed increased proliferation and invasion,whereas cells transfected with siCaveolin-1 showed decreased proliferation and invasion.Patients with HCC with higher miR-124 and lower caveolin-1 levels lived longer than patients with lower miR-124 and higher caveolin-1 levels.Conclusions:MiR-124 mediated the proliferation and invasion of HCC by targeting caveolin-1.