微小核糖核酸-205靶向调控血管内皮生长因子-A对肝癌细胞的影响

The inhibition effect of microRNA-205 on hepatocellular carcinoma cells through targeting regulation of the vascular endothelial growth factor-A

目的探讨微小核糖核酸(miR)-205在肝细胞癌(HCC)中的作用及其与肝癌进展的关系,以及miR-205是否通过靶向血管内皮细胞生长因子(VEGF)-A影响肝癌细胞增殖。方法采用逆转录-定量聚合酶链反应(RT-qPCR)检测miR-205在HCC、相邻正常肝组织和肝癌细胞株HepG2、HuH-7、SMMC-7721、BEL-7402、人正常肝细胞株L02中的表达。RT-qPCR检测转染miR-205激动剂和空白对照的HepG2和HuH-7细胞中miR-205表达。Transwell和溴化噻唑蓝四氮唑(MTT)法检测转染后肝癌细胞增殖、迁移和侵袭。双荧光素酶报告基因检测VEGF-A是否为miR-205直接作用靶标。VEGF-A siRNA敲低分析VEGF-A表达是否为miR-205调控HCC细胞增殖、迁移和侵袭的关键介质。结果miR-205在HCC组织和肝癌细胞株中表达下调。miR-205过表达可抑制肝癌细胞增殖、迁移和侵袭。VEGF-A是HCC中miR-205直接作用靶标,miR-205通过靶向VEGF-A抑制肝癌细胞增殖、迁移和侵袭。结论miR-205可通过下调VEGF-A表达抑制HCC生长、迁移和侵袭,这可能是未来HCC治疗的潜在靶点。

Objective To explore of the inhibition effect of mirRNA-205(miR-205) on hepatocellular carcinoma(HCC), to analyze the relationship between miR-205 and HCC progression, and to clarify whether the inhibition effect of miR-205 on HCC is achieved by targeting regulation of vascular endothelial growth factor-A(VEGF-A). Methods Reverse transcription-quantitative polymerase chain reaction(RT-qPCR) was used to detect the expressions of miR-205 in HCC, adjacent normal liver tissues, hepatocellular carcinoma cell lines of HepG2, HuH-7, SMMC-7721, BEL-7402, and human normal liver cell line L02. RT-qPCR was adopted to test the expressions of miR-205 in HepG2 and HuH-7 cells transfected with miR-205 agonist a nd blank control. Transwell and MTT methods were employed to test the proliferation, migration and invasion of HCC cells after transfection. Luciferase assay system was adopted to determine whether VEGF-A was a direct target of miR-205. VEGF-A siRNA knock down analysis was conducted to clarify whether the expression of VEGF-A was a key mediator in regulating the proliferation, migration and invasion of HCC cells.Results The expression of miR-205 was down-regulated in both HCC tissues and HCC cell lines. The over-expression of miR-205 could inhibit the proliferation, migration and invasion of HCC cells. VEGF-A was a direct target of miR-205 in HCC. By down-regulating the expression of VEGF-A, miR-205 exerted its inhibition effect on the proliferation, migration and invasion of HCC cells. Conclusion Through down-regulating the expression of VEGF-A, miR-205 achieves its inhibition effect on the proliferation, migration and invasion of HCC cells, which may be a potential target for the treatment of HCC in the future.