微小RNA-5586-5p在肝细胞癌中的表达及其对肝细胞癌侵袭、迁移的影响

目的检测肝细胞癌中miRNA-5586-5p的表达情况,探究其与肝细胞癌临床病理特征和肝细胞癌病人预后的相关性,进而探究其对肝细胞癌细胞侵袭和迁移的影响。方法选取2010年1月—2013年6月西安交通大学第一附属医院进行根治性肝癌切除术的肝细胞癌患者100例。采用Real-time PCR法检测肝细胞癌组织和癌旁组织中miRNA-5586-5p的表达水平;采用合理统计学方法进行miRNA-5586-5p的表达水平与肝细胞癌临床病理特征的相关性分析及对肝细胞癌患者预后的影响。采用Real-time PCR法检测肝细胞癌细胞系中miRNA-5586-5p的表达水平,继而通过Transwell小室实验研究miRNA-5586-5p对肝细胞癌细胞侵袭和迁移能力的影响。采用Real-time PCR、Western blotting以及免疫组织化学检测miRNA-5586-5p下游潜在靶点肝细胞癌水通道蛋白9(AQP9)的m RNA和蛋白表达水平,利用Western blotting证实miR-5586-5p下调AQP9并通过miRNA-5586-5p-AQP9-AKT通路发挥促癌作用。结果 miR-5586-5p在肝细胞癌组织中的表达水平较癌旁组织显著上调(P0.01)。miRNA-5586-5p的高表达与肿瘤数量、血管侵犯、Edmondson分级和TNM分期显著相关(P0.05、0.01)。与mi RNA-5586-5p高表达者比较,miR-5586-5p低表达者的生存时间更长(P0.01)。miR-5586-5p在肝细胞癌细胞系中的表达均明显高于正常肝细胞(P0.05),经Transwell小室实验证实miR-5586-5p具有增强肝细胞癌细胞的侵袭、迁移的作用。miR-5586-5p可下调AQP9的蛋白表达量;且miR-5586-5p与AQP9蛋白表达呈负相关。结论 miR-5586-5p在肝细胞癌中为高表达,并显著促进肝细胞癌的侵袭、迁移能力,miR-5586-5p与AQP9的蛋白水平呈负相关,并且能够抑制AQP9表达和通过miR-5586-5p-AQP9-AKT通路促进肝细胞癌细胞的侵袭与迁移能力。

Expression of micro RNA-5586-5p in hepatocellular carcinoma and its effect on invasion and migration of hepatocellular carcinoma

Objective To detect the expression of microRNA-5586-5p in hepatocellular carcinoma, explore its correlation with clinicopathological characteristics and prognosis of hepatocellular carcinoma patients, and further explore its influence on invasion and migration of hepatocellular carcinoma cells. Methods Patients (100 cases) with hepatocellular carcinoma in The First Affiliated Hospital of Xi''an Jiaotong University from January 2010 to June 2013 were selected. Detection of the microRNA-5586-5p expression in hepatocellular carcinoma and its adjacent tissues were carried on by Real-time PCR. The correlation between the expression of microRNA-5586-5p and the clinicopathological features of hepatocellular carcinoma, and the prognosis of patients with hepatocellular carcinoma were analyzed by rational statistical methods. Real-time PCR was used to detect the expression levels of miRNA-5586-5p in hepatocellular carcinoma cell lines, and then the effect of miRNA-5586-5p on invasion and migration of hepatocellular carcinoma cells was studied by Transwell assay. Real-time PCR, Western blotting and immunohistochemistry were used to detect the mRNA and protein expression of the downstream potential target of miRNA-5586-5p in AQP9 of hepatocellular carcinoma, and Western blotting was used to confirm that miRNA-5586-5p down-regulated AQP9 and played a pro-cancer role through the miRNA-5586-5p-AQP9- AKT pathway. Results The expression level of miRNA-5586-5p in hepatocellular carcinoma tissues was significantly higher than that in adjacent tissues (P<0.01). The high expression of miRNA-5586-5p was significantly correlated with the number of tumors, vascular invasion, Edmondson grade and TNM grade (P<0.05, 0.01). Compared with miRNA-5586-5p high expression group, the miRNA-5586-5p low expression group had longer survival time (P<0.01). The expression of miRNA-5586-5p in hepatocellular carcinoma cell lines was significantly higher than that in normal hepatocytes (P<0.05). Transwell experiments showed that miRNA-5586-5p could enhance the invasion and migration of hepatocellular carcinoma cells. miRNA -5586-5p could down-regulate the expression of AQP9 protein, and the expression of AQP9 protein was negatively correlated with that of miRNA -5586-5p. Conclusion miR-5586-5p is highly expressed in hepatocellular carcinoma and significantly promotes the invasion and migration of hepatocellular carcinoma. And miR-5586-5p is negatively correlated with AQP9 protein level, and can inhibit the expression of AQP9 and promote the invasion and migration of hepatocellular carcinoma cells through the miR-5586-5p-AQP9-AKT pathway.