西妥昔单抗联合塞来昔布对肺腺癌细胞KDR和AQP1表达的影响

Effects of Cetuximab Combined with Celecoxib on Apoptosis and KDR and AQP1 Expression in Lung Cancer

背景与目的新辅助化疗是肺癌治疗的新进展,西妥昔单和合塞来昔布均是近年来新辅助化疗的常用药物,本研究旨在探讨表皮生长因子受体(epithelial growth factor receptor,EGFR)单克隆抗体西妥昔单抗(Cetuximab)联合环氧合酶2(cyclooxygenase-2,COX-2)抑制剂塞来昔布对肺腺癌A549细胞株凋亡和KDR、AQP1表达的影响。方法 A549细胞培养于RPMI-1640培养液中,实验分为正常对照组、西妥昔单抗1 nmol/L组、塞来昔布25μmol/L组、西妥昔单抗1 nmol/L+塞来昔布25μmol/L组。药物干预细胞48 h后,采用四甲基偶氮唑盐比色法测定细胞抑制率;采用流式细胞术测定细胞周期分布的变化;用Transwell小室法检测药物处理前后细胞侵袭力的变化。采用RTPCR、Western blot检测KDR、AQP1基因和蛋白的表达情况。结果西妥昔单抗和塞来昔布对A549细胞增殖有明显的抑制作用,两制剂联合作用时抑制作用更强(P<0.01)。西妥昔单抗和塞来昔布均可诱导细胞凋亡,联合作用后细胞凋亡率更高(P<0.01);联合用药与单独用药相比,可使细胞发生明显的G1期阻滞(P<0.01),进一步下调了KDR、AQP1基因和蛋白的表达(P<0.05),细胞侵袭力明显降低。结论西妥昔单抗和塞来昔布联合应用具有协同作用,可进一步抑制细胞的生长和迁移,下调了KDR、AQP1基因和蛋白的表达提示两药联合使用临床肺癌治疗可能具有很大的潜力。

Background and objective Neoadjuvant chemotherapy is a new development in the treatment of lung cancer. In recent years, cetuximab and celecoxib have been commonly used in this procedure. This study aims to explore the effect of cetuximab combined with celecoxib on apoptosis and KDR and AQP1 expression in lung cancer A549 cells. Method The cells were cultured in RPMI-1640 and then divided into four groups： control group, 1 nmol/L cetuximab group, 25 μmol/ L celecoxib group, and 1 nmol/L cetuximab＋25 μmol/L celecoxib group. The treatment time was 48 h. The mRNA and protein expression levels of KDR and AQP1 were detected by RT-PCR and Western blot, respectively. The apoptosis, proliferation, and invasive ability ofA.549 cells before and after transfection were examined using flow cytometry, MTT, and transwell meth- ods. Results Cetuximab and celecoxib inhibited the growth ofA549 cells in a dose-dependent manner. Their combination produced a greater growth inhibition than when either was used alone （P〈0.01）. Cetuximab and celecoxib both induced the apoptosis ofA549 cells, and their combination produced a higher apoptosis rate （P〈0.01）. Cetuximab in combination with ce- lecoxib also induced G1 phase arrest and downregulated the expression of KDR and AQ_P 1 in A549 cells （P〈0.05）. As a result, the invasion ability of the A549 cells was significantly decreased. Conclusion Cetuximab in combination with celecoxib can synergistically inhibit the growth ofA.549 cells and downregulate the expression of KDR and AQP1 in A.549 cells. The combi- nation of cetuximab and celecoxib is a potential strategy for lung cancer therapy.