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miR-605-3p通过靶向ZIK1抑制肝细胞癌迁移和侵袭
作者:胡艳  黄智平  周维  张红卫  俞雷 
单位:上海市第七人民医院肿瘤科  上海 200137 广州军医广州总医院肝脏外科  广东 广州 510010 
关键词: 肝细胞/病理学 微RNAs 细胞运动 肿瘤侵润 生物标记 肿瘤 肿瘤细胞 培养的 聚合酶链反应 
DOI:10.13267/j.cnki.syzlzz.2020.01.006
出版年,卷(期):页码:2020,35(1):30-36
摘要:

目的 探讨miR-605-3p对肝细胞癌细胞迁移和侵袭能力的影响及其相关机制。方法 采用定量PCR检测80例肝癌组织及其配对癌旁组织以及肝癌细胞株(Huh7、HCC-LM3、Hep3B以及SMMC-7721)和肝细胞株(THLE-3)中miR-605-3p的表达水平。生物信息学分析预测miR-605-3p结合的靶蛋白,Western blot检验ZIK1蛋白的表达情况,荧光素酶报告验证miR-605-3p与ZIK1之间的相互关联。Kaplan-Meier曲线和Log-rank分析方法比较miR-605-3p高、低表达组无进展生存期和总生存期。将SMMC-772细胞分为模拟物转染组、抑制物转染组、模拟物对照组和抑制物对照组。CCK8增殖实验检测细胞增殖能力,细胞划痕实验和transwell侵袭实验检测细胞迁移和侵袭。结果 miR-605-3p在肝癌组织中表达下调(P<0.05)。以miR-605-3p在肝癌组织中位表达分为高表达和低表达组,miR-605-3p在肝癌组织低表达患者5年总生存率(21.6% vs 48.3%,P<0.05)和无进展生存率均低于高表达患者(9.5% vs 30.8%,P<0.05)。miR-605-3p抑制剂可以增强肝癌细胞迁移和侵袭能力(均P<0.05)。生物信息学分析证实,ZIK1蛋白是miR-605-3p肝癌细胞中的直接作用靶点。结论 miR-605-3p在肝癌发生和发展过程中发挥作用,通过调控ZIK1的表达影响肝癌细胞的迁移和侵袭能力。

Objective To investigate the effect of microRNA-605-3p (miR-605-3p) on the migration and invasion of hepatocellular carcinoma (HCC) cells and its related mechanism. Methods Quantitative PCR was used to detect the expression of miR-605-3p in 80 HCC tissue samples and their paracancerous tissues as well as HCC cell lines (Huh7,HCC-LM3,Hep3B and SMMC-7721) and hepatocyte cell line (THLE-3). Bioinformatics analysis predicted the target proteins binding to miR-605-3p,Western blot analysis was used to detect ZIK1 protein expression,and the luciferase report verified the correlation between miR-605-3p and ZIK1 protein. Kaplan-Meier curve and Log-rank analysis methods were used to compare the difference of progression-free survival and overall survival between the high and low expression groups of miR-605-3p. SMMC-772 cells were divided into the analogue transfection group,inhibitor transfection group,analogue negative control group,and inhibitor negative control group. CCK8 proliferation test was used to detect cell proliferation,and cell scratch assay and transwell invasion assay were used to detect cell migration and invasion. Results The expression of miR-605-3p was down-regulated in HCC tissues (P<0.05). The median expression of miR-605-3p in HCC tissues was used to divide patients into the high-expression and low-expression groups. The 5-year overall survival rate (21.6% vs 48.3%,P<0.05) and progression-free survival rate of the patients with low expression of miR-605-3p in HCC tissues were lower than those with high expression (9.5% vs 30.8%,P<0.05). miR-605-3p inhibitors could enhance the migration and invasion ability of HCC cells (all P<0.05). Bioinformatics analysis confirmed that ZIK1 protein was the direct target of miR-605-3p in HCC cells. Conclusions miR-605-3p plays an important role in the occurrence and development of HCC,and affects the migration and invasion of HCC cells by regulating the expression of ZIK1.

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