miR-190通过调控细胞因子信号转导抑制因子5促进肝细胞癌发生的机制

目的 探讨微RNA-190（miR-190）通过调控细胞因子信号转导抑制因子5（SOCS5）介导肝细胞癌（HCC）发生的机制。方法 选取84例HCC患者的肝癌组织及癌旁组织,通过实时荧光定量PCR（RT-PCR）检测miR-190表达水平,并分析其与患者临床特征和预后的相关性。将miR-190模拟物/抑制物或SOCS5 siRNA（siSOCS5）转染至SNU398和HepG2细胞中,CCK-8检测细胞的增殖,Transwell对细胞的侵袭与迁移进行检测,RT-PCR和蛋白免疫印迹法检测SOCS5基因和蛋白的表达水平,双荧光素酶测定验证miR-190与SOCS5的靶向关系。通过异种移植肿瘤验证SOCS5体内抑癌作用。结果 HCC中miR-190水平上调,且miR-190高表达与肿瘤大小、淋巴结转移、临床分期、预后不良有关（P均 < 0.05）。CCK-8和Transwell结果表明,过度表达的miR-190可促进细胞增殖、侵袭及迁移。RT-PCR、蛋白免疫印迹法和双荧光素酶测定结果证实了SOCS5是miR-190的真正靶点。此外,SOCS5在体内外均有促进HCC发展的作用。结论 miR-190可通过靶向SOCS5促进HCC的发生发展,抑制miR-190在恢复HCC中SOCS5水平方面有潜在治疗用途。

The mechanism of miR-190 promoting the incidence of hepatocellular carcinoma by regulating the suppressor of cytokine signaling-5

Objective To investigate the mechanism of microRNA-190 （miR-190） mediating the incidence of hepatocellular carcinoma （HCC） through regulating the suppressor of cytokine signaling-5 （SOCS5）. Methods The liver cancer tissues and adjacent tissues were collected from 84 HCC patients. The expression level of miR-190 was detected by real-time fluorescent quantitative PCR（RT-PCR）, and its correlation with clinical characteristics and prognosis of the patients was analyzed. miR-190 mimic/inhibitor or SOCS5 siRNA （siSOCS5） were transfected into the SNU398 and HepG2 cells. Cell proliferation was detected by CCK-8 assay. Cell invasion and migration were detected by Transwell assay. The expression levels of SOCS5 mRNA and protein were determined by RT-PCR and Western blot. The targeted relationship between miR-190 and SOCS5 was verified by dual-luciferase assay. Tumor xenotransplantation was performed to verify the tumor-suppressing effect of SOCS5 in vivo. Results The expression level of miR-190 in HCC was up-regulated, and high expression of miR-190 was associated with tumor size, lymph node metastasis, clinical stage and poor prognosis（all P < 0.05）.The results of CCK-8 and Transwell assay showed that over-expression of miR-190 could promote cell proliferation, invasion and migration. RT-PCR, Western blot and dual-luciferase assay confirmed that SOCS5 was the true target of miR-190. In addition, SOCS5 could promote the development of HCC both in vivo and in vitro. Conclusion The miR-190 can promote the incidence and development of HCC by targeting SOCS5, indicating that suppressing miR-190 possesses the potential therapeutic value in restoring the SOCS5 level in HCC.