趋化因子受体CXCR4在肝细胞癌表达及肺转移中作用的研究

Role of chemokine receptor CXCR4 in lung metastasis of human hepatoceilular carcinoma

目的以人肝细胞癌高、低肺转移潜能细胞株MHCC97H、MHCC97-L为研究对象，研究趋化因子受体CXCR4在肝细胞癌肺转移过程中的作用和意义。方法RT-PCR和Western blotting比较MHCC97H、MHCC97-L细胞株CXCR4 mRNA及蛋白质水平的表达。CXCR4配体SDF-1α及肺提取物趋化实验和抗体抑制实验观察SDF-1α和裸鼠肺组织匀浆液对MHCC97-H的趋化迁移作用。结果趋化因子受体CXCR4在MHCC97-H细胞株表达低于低肺转移潜能细胞株MHCC97-L，蛋白质水平与mRNA水平一致。SDF-1α对MHCC97-H趋化实验表明在1～100ng／ml浓度范围内均有趋化作用，在浓度为50ng／L时趋化作用最显著（P〈0．05）。肺提取物亦发现对MHCC97-H有趋化作用，作用强于MHCC97-L及DMEM组（P〈0．05）。但加入CXCR4抗体后其趋化作用并未明显下调。结论CXCR4在肝癌组织及细胞表达并可能参与肝癌肺转移，但并非肝癌细胞趋化转移的主要受体分子。

Objective To study the role of chemokine receptor CXCR4 in lung metastasis of hepatocellular carcinoma （HCC） through human HCC cell line MHCC97-H and MHCC97-L with different lung metastasis potentials. Methods The mRNA and protein expression of CXCR4 between MHCC97-H and MHCC97-L by were determined by RTPCR and Western blotting respectively. Chemotaxis and antibody inhibition assays were used to observe the chemotaxis effects of SDF la, specific ligant of CXCR4, and lung crude extraction from nude mice on MHCC97-H. Results Chemokine receptor CXCR4 in MHCC97-H was expressed lower than that in MHCC97-L, and protein level was in consistent with the mRNA level. MHCC97-H could respond to SDF-1α between 1 ng/ml to100 ng/ml, particularly at 50 ng/L（P〈0.05）. Lung extraction could have chemotaxis to MHCC97-H, compared with other groups （P,0.05）. However, the chemotaxis effect decreased little when being added with CXCR4 antibody. Conclusion Chemokine receptor CXCR4 may participate in the lung metastasis of HCC but is not the major chemokine receptor involved.