microRNA-19对肝癌细胞系迁移与侵袭能力的影响

目的 比较不同转移潜能肝癌细胞系中microRNA-19（mir-19）表达变化,探讨mir-19对不同转移潜能肝癌细胞迁移和侵袭能力的影响.方法 应用TaqMan MGB探针法定量分析mir-19在高转移潜能肝癌细胞系（MHCC97H、LM3）和低转移潜能肝癌细胞系（HepG2、7402）的表达差异.利用转染试剂将人工合成的mir-19的前体转染高转移潜能细胞系,人工合成的mir-19抑制剂转染低转移潜能细胞系.采用细胞计数试剂盒-8（Cell Counting Kit-8,CCK-8）检测mir-19对细胞增殖的影响,Transwell小室检测mir-19对细胞的迁移和侵袭能力的影响.结果 在不同转移潜能的细胞系中,mir-19表达水平在高转移潜能细胞系较低转移潜能细胞系表达下调（P〈0.05）.功能实验显示mir-19可抑制肝癌细胞的侵袭和增殖（P〈0.05）,利用生物信息学预测mir-19的靶基因发现,许多癌基因3＇UTR包含与mir-19的互补序列.结论 mir-19可能部分通过调节CREB3L4、PDE4A、JAM2、RUNX2等的表达来抑制肝癌细胞的增殖和侵袭转移.

Effect of microRNA-19 on Migration and Invasion of Hepatoma Carcinoma Cell

Objiective To analyze the differential expressions of mir-19 in different metastasis potential HCC cell line by quantitative real time PCR.We carried out further analyses mir-19 to determine whether it is functionally linked to migration and invasion of hepatoma carcinoma cell. Methods A qRT-PCR method was established and using special looped-primers and TaqMan MGB quencher to detect the relative expression level of mir-19 and differernt metasitasis potential HCC cell line.Precursor or inhibitor of mir-19 was transfected into HCC cells with transfection agent.Cell counting kit-8 and transwell assay was carried out in transfected HCC cell lines to assess its influence on cell growth,migration,and invasion. Results mir-19 were significantly expressed between high metastasis cell lines （MHCC97H and LM3） and low metastasis cell lines（HepG2 and 7402）,which showed lower expression in the former（P〈O.OS）.Functional studies indicated that mir-19 significantly inhibited HCC cell migration and cell growth.In-silico analysis revealed CREB3L4,PDE4A,JAM2,RUNX2,et al as potential targets of mir-19. Conclusion mir-19 may suppress the migration and invasion of the hepatoma carcinoma cell.HCC partially through targeting CREB3L4,PDE4A,JAM2,RUNX2 and et al.