埃罗替尼和塞来昔布协同抑制胆管癌细胞生长

Synergistic inhibitory effect of erlotinib and celecoxib on the growth of cholangiocarcinoma cells

目的:观察表皮生长因子受体(EGFR)酪氨酸激酶抑制剂埃罗替尼(erlotinib)与环氧合酶(COX-2)抑制剂塞来昔布(celecoxib)对胆管癌细胞生长的协同抑制作用。方法:采用甲基噻唑基四唑(MTT)比色法检测胆管癌细胞株QBC939增殖,流式细胞术检测用药前、后细胞凋亡和细胞周期的变化,逆转录聚合酶链反应(RT-PCR)、Western印迹方法观察用药前、后细胞中COX-2、EGFR下游信号及凋亡、细胞周期相关基因蛋白表达的变化。结果:QBC939细胞表达COX-2 mRNA和EGFR mRNA及相应蛋白。埃罗替尼、塞来昔布抑制QBC939细胞增殖,促进细胞凋亡,诱导细胞周期阻滞于G0/G1期。同时,塞来昔布增强了埃罗替尼抑制细胞生长、诱导细胞凋亡的作用。联合用药显著降低p-MAPK、p-Akt及PGE2表达。结论:塞来昔布和埃罗替尼均可抑制胆管癌细胞的生长,而联合用药具有协同作用,能同时阻断EGFR和COX-2信号通路。在胆管癌治疗中具有一定应用前景。

Objective Epidermal growth factor receptor（EGFR） and cyclooxygenase-2（COX-2） contribute to the development of cholangiocarcinoma.Blocking simultaneously both EGFR and COX-2 mediated pathways may be an efficient means of inhibiting cancer cell growth in cholangiocarcinoma.Methods A combination of EGFR-selective tyrosine kinase inhibitor（TKI） erlotinib and a COX-2 inhibitor（COX-2I） celecoxib was studied for their effects on the cell growth,cell cycle progression,and apoptosis in cholangiocarcinoma cell line by cell growth assay and flow cytometric analysis.Primary and secondary targets of EGFR TKIs and COX-2 were also examined using RT-PCR and immunoblotting assay after the combined treatment.Results The combination of erlotinib with celecoxib synergistically inhibited the growth of the QBC939 cell line,significantly induced G0/G1 arrest and apoptosis.Furthermore,the combination as compared with the effect of single agents showed strong reductions of p-MAPK,p-Akt and PGE2 of the cholangiocarcinoma cell line.Erlotinib inhibited the expression of COX-2 protein,whereas celecoxib mainly blocked the production of prostaglandin E2 in QBC939 cells.Conclusions The results suggest that cell growth inhibition induced by a combination of EGFR TKIs and COX-2 is mediated through blocking simultaneously the EGFR and COX-2 pathways.This combination holds a great potential for the treatment of cholangiocarcinoma.