摘要
目的观察趋化因子受体（CCR1 CCR7 CXCR3 CXCR4）在人肝癌细胞系（Hep3B HepG2 HLE和HLF）的表达，并探讨其作用机制。方法采用逆转录-聚合酶链反应（RT—PCR）、细胞免疫组织化学、流式细胞仪测定趋化因子受体在肝癌细胞表面的表达，通过肝癌细胞Hep3B体外侵袭实验即重组大鼠巨噬细胞激动蛋白（CCL3）-1对Hep3B细胞穿透人工重组基底膜能力的影响来检测趋化因子受体在肝癌侵袭转移时所起的作用，并通过激光共聚焦显微镜观察趋化因子受体在肝癌细胞侵袭转移过程中的作用机制。结果通过RT—PCR，流式细胞术可检测到CCR1 mRNA及蛋白在肝癌细胞中表达，免疫组织化学显示CCR1在肝癌细胞包膜和胞质内都有表达。细胞侵袭实验提示Hep3B在CCL3的刺激下，实验组Hep-3B细胞通过基底膜的数量（213±12）多于对照组细胞通过基底膜的数量（102±11），差异有统计学意义（P〈0．01）。运用激光共聚焦检测到实验组胞内钙离子的浓度增加到。结论肝癌细胞系（Hep3 BHepG2 HLE和HLF）的表面的表达CCR1，且CCR1在肝癌细胞侵袭转移中发挥重要的作用，其作用机制与细胞内钙离子浓度有很关。
Objective To investigate the expression and mechanism of chemokine receptors （ CCR1, CCR7, CXCR3, CXCR4） in hepatoma carcinoma cells （ Hep3 B, HepG2, HLE and HLF）. Methods RT-PCR immunohistochemistry and flow cytometric were used to detect the expression of CCR1 chemokine receptor in hepatoma carcinoma cells. Invasion of Hep3B was analyzed with basal membrane by CCL3 and action mechanism of chemokine receptor was analyzed in hepatoma carcinoma cells by confocal microscopy. Results CCR1 mRNA and protein were detectable in hepatoma carcinoma cells. Immunohistochemistry revealed that CCR1 protein was located on membrane and in cytoplasm of hepatoma carcinoma cells. The number of hepatoma carcinoma cells that went through basal membrane was significantly more in experiment group （213 ± 12） than in control group （ 102 ± 11 ） （P 〈 0.01 ）. The density of the calcium ion was increased in hepatoma carcinoma cells in experiment group. Conclusion Hepatoma carcinoma cells express CCR1 chemokine recepter, and CCR1 plays an important role in the process of metastasis and invasion of hepatoma carcinoma cells. The density of the calcium ion has a great relationship with the mechanism of chemokine receptor in hepatoma carcinoma.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2008年第11期1366-1368,F0003,共4页
Chinese Journal of Experimental Surgery
关键词
癌
肝细胞
趋化因子受体
Carcinoma,hepatocellular
Chemokine receptor
