血管内皮生长因子受体-1促进肝癌细胞侵袭和转移的初步研究

Vascular endothelial growth factor receptor-1 promotes migration and invasion of hepatocellular carcinoma cell lines

目的检测不同转移潜能的肝癌细胞中有无血管内皮生长因子受体-1(VEGFR-1)及其配体(VEGF)的表达以探讨VEGFR-1在肝癌侵袭转移中的作用。方法分别用RT-PCR、ELISA和/或Western blot法检测四种肝癌细胞株MHCC97-H、MHCC97-L、SMMC 7721和HepG2有无VEGFR-1 mRNA和蛋白表达,用VEGFR-1的特异性配体VEGF-B处理MHCC97-H细胞以研究VEGFR-1活化对肝癌细胞迁移、侵袭和增殖等的影响。结果MHCC97-H、MHCC97-L和SMMC 7721有VEGFR-1 mRNA和蛋白表达,但4种肝癌细胞都有VEGFR-1的配体VEGF-A和VEGF-B的表达;VEGFR-1活化能促进肝癌细胞MHCC97-H侵袭和迁移;VEGFR-1依赖的侵袭和迁移能被VEGFR-1的中和抗体18F1阻断,VEGFR-1活化不能促进细胞增殖和存活。结论VEGFR-1及其配体的表达与肝癌细胞的侵袭转移有关,肝癌中的VEGF过表达可以自分泌的方式激活表达VEGFR-1的肝癌细胞,VEGFR-1及其配体可能是防治原发性肝癌侵袭转移的新靶点。

Objectives To investigate the role of vascular endothelial growth factor receptor- 1 （VEGFR- 1 ） in invasion and metastasis of hepatocellular carcinoma cell lines by testing the expression of VEGFR-1 and its ligands （VEGFs）. Methods The mRNA and protein expressions of VEGFR-1 and/or VEGFs in MHCC97-H, MHCC97-L, SMMC7721, and HepG 2 cells lines were tested by reverse-transcriptasepolymerase chain reaction （RT-PCR）, enzyme linked immunosorbent assay （ELISA）, and Western blot （WB） analysis. The function of VEGFR-1 was evaluated by treating MHCC97-H cell lines with VEGF-B （a selective ligand for VEGFR-1 ） and/or a specific anti-VEGFR-1 antibody. Results MHCC97-H, MHCC97-L, and SMMC7721 cell lines expressed VEGFR-1 mRNA and protein, as well as the VEGFR-1 ligands VEGF-A and VEGF-B. However, HepG 2 only expressed the VEGF-A and VEGF-B but no VEGFR-1 mRNA and protein. VEGFR-1 activation by VEGF-B was promoted migration in MHCC97-H cell lines. MHCC97-H cells also demonstrated enhanced matrigel invasion after VEGFR-1 stimulation. VEGFR-l-dependent migration and invasion were blocked by the VEGFR-1 neutralizing antibody 18F1. VEGFR-1 activation did not appear to enhance cell proliferation. Conclusion The study has demonstrated the relation between the expressions of VEGFR-1 and its ligands and the migration and invasion in hepatocellular carcinoma. Overexpression of VEGF in hepatocellular carcinoma may activate tumor cells bearing VEGFR-1 via an autocrine pathway. VEGFR-1 and its ligands may be new targets for preventing and curing the metastatis and invasion.