二甲双胍联合塞来昔布对胰腺癌细胞增殖及Caspase-3活性的影响

The Effects of Metformin Combined with Celecoxib on the Proliferation and Caspase-3 Activity in Pancreatic Cancer Cells

目的:探讨二甲双胍和塞来昔布单独或联合应用对胰腺癌细胞增殖和Caspase-3活性的影响。方法:体外培养人胰腺癌BxPC-3和As PC-1细胞,用四甲基偶氮唑盐(MTT)法检测不同浓度二甲双胍和塞来细胞对胰腺癌细胞存活率的影响。联合实验分4组:对照组、二甲双胍组(MET,15 mmol/L)、塞来昔布组(CEL,100μmmol/L),二甲双胍和塞来昔布联合组(MET 15 mmol/L+CEL100μmmol/L),孵育48 h,用MTT检测细胞存活率,用Caspase-3比色测定试剂盒测定Caspase-3活性。结果:二甲双胍和塞来昔布单药均可以时间和剂量依赖性方式降低胰腺癌Bx PC-3和As PC-1细胞的生存率。两种细胞系的各加药组细胞存活率与对照组比较差异均存在统计学意义(P<0.01),联合实验组的细胞存活率明显低于单独用药组(P<0.01)。二甲双胍和塞来昔布单药处理的胰腺癌Bx PC-3和As PC-1细胞Caspase-3活性均显著高于对照组(P<0.01),二甲双胍和塞来昔布联合实验组Caspase-3活性均明显高于单药处理组和对照组(P<0.01),但二甲双胍组和塞来昔布组之间的Caspase-3活性比较差异无统计学意义(P>0.05)。结论:二甲双胍和塞来昔布联合作用可协同抑制胰腺癌细胞的增殖,并通过激活Caspase-3活性促进胰腺癌细胞的凋亡,其联合应用可能成为胰腺癌药物治疗的有效策略。

Objective： To study the effects of metformin and celecoxib alone or in combination on the cell viability and Caspase-3 activity in the pancreatic cancer BxPC3 and AsPC-1 cells. Methods： Pancreatic cancer cell line BxPC-3 and AsPC-1 were cultured and treated with different concentrations of mefformin or celecoxib for 24, 48 and 72 h, the cell viability was then measured and calculated by the MTT assay. In the combination experiments, each cell line was divided into 4 groups： the untreated control group, metformin group alone （treated with 15 mmol/L metformin）, celecoxib group alone （treated with 100 μmmol/L celecoxib）, the combination group（met- formin 15 mmol/L＋ celecoxib 100 μmmol/L ）. After treated for 48 h, MTT assay was used for the detection of viability and Caspase-3 activity. Results： The viability of BxPC-3 and AsPC-1 cell were inhibited by metformin or celecoxib in a time- and dose-dependent man ner. Compared with the untreated control group, the cell viability of all the drug treated groups were significantly decreased, while the Caspase-3 enzyme activity were increased in both BxPC-3 and AsPC-1 cells （P〈0.01）. Compared to the untreated control or individual drug, the combination of metforrnin and celecoxib significantly decreased cell viability and increased caspase-3 enzyme activity of both cells （P〈0.01）. There were no significant different in caspase-3 enzyme activity between metformin group and celecoxib group（P〉0.05）. Conclusion： Combination ofmetformin and celecoxib could synergistically inhibite the cell proliferation and increased caspase-3 enzyme activity, thus induced apoptosis in the pancreatic cancer cell, which indicated that combination of metformin and celecoxib might be a promising remedy in treatment of pancreatic cancer.