培美曲塞联合奥沙利铂对人胃癌BGC-823细胞生长的影响

Effect of pemetrexed combined with oxaliplatin on the growth of human gastric cancer cell BGC-823

目的:探讨培美曲塞与奥沙利铂对人胃癌细胞BGC-823的生长抑制作用以及可能的作用机制。方法:采用不同浓度培美曲塞和奥沙利铂单独或联合作用于BGC-823细胞。应用MTT法检测BGC-823细胞生长抑制率,FCM检测细胞周期的变化,蛋白质印迹法检测增殖细胞核抗原(proliferating cell nuclear antigen,PCNA)和细胞周期蛋白D1(cell cycle D1,cyclinD1)的表达水平。结果:培美曲塞和奥沙利铂均可抑制BGC-823细胞的生长(P<0.05),且呈时间和剂量依赖性。这两种药物均可明显增加G1期细胞比例,降低S期细胞比例。蛋白质印迹法检测结果显示,cyclinD1和PCNA表达量均明显降低,其中培美曲塞和奥沙利铂联合处理具有协同效应,培美曲塞(80μg/mL)处理24h后序贯奥沙利铂(40μg/mL)处理24h组BGC-823细胞的生长抑制率明显高于双药联合处理组以及奥沙利铂(40μg/mL)处理24h后序贯培美曲塞(80μg/mL)处理24h组(P<0.05)。结论:培美曲塞和奥沙利铂均可抑制胃癌细胞BGC-823的生长,细胞被阻滞于G1期。在双药联合处理中,以培美曲塞序贯奥沙利铂对胃癌细胞的生长抑制最强。

Objective： To investigate the growth inhibition effects of pemetrexed and oxaliplatin on human gastric cancer cell line BGC-823 in vitro, and to explore the possible mechanism. Methods： The BGC-823 cells were treated with different concentrations of pemetrexed or oxaliplatin alone or in combination. The growth inhibition rate of BGC-823 cells was detected by MTT method, and the change of cell cycle distribution was analyzed by flow cytometry （FCM）. The expressions levels of proliferating cell nuclear antigen （PCNA） and cell cycle D1 （cyclin D1） proteins in BGC-823 cells were examined by Western blotting. Results： The growth of BGC-823 cells was inhibited by pemetrexed or oxaliplatin in a dose- and time-dependent manner （P〈0.05）. The percentage of cells in GT phase was increased while which in S phase was decreased by the treatment of pemetrexed or oxaliplatin. The expression levels of PCNA and cyclin D1 proteins were significantly decreased, and the combination regimen of pemetrexed and oxaliplatin exerted a synergistic effect. The growth inhibition rate of BGC-823 cells treated with pemetrexed （80 μg/mL） for 24 h followed by oxaliplatin （40 μg/mL） for 24 h was significantly decreased as compared with those of the cells treated with pemetrexed （80 μg/mL） combined with oxaliplatin （40 μg/mL） at the same time or treated with oxaliplatin （40 μg/mL） for 24 h followed by pemetrexed （80 μg/mL） for 24 h （P〈0.05）. Conclusion： Pemetrexed and oxaliplatin can both inhibit the proliferation of gastric cancer BGC-823 cells and arrest the cells at G1 phase. The sequential regimen of pemetrexed followed by oxaliplatin may have the strongest growth-inhibitory effect on gastric cancer cells among different doublet regimens.