阿司匹林与氟尿嘧啶协同抑制结肠癌细胞生长增殖的机制研究

Synergistic anti-proliferation effect of aspirin and 5-fluorouracil on colon cancer cells and its mechanism

目的:检测阿司匹林与氟尿嘧啶协同抑制结肠癌细胞生长增殖的能力及相关作用机制。方法:将体外培养的结肠癌细胞分为4组:空白对照组、阿司匹林组、氟尿嘧啶组和阿司匹林+氟尿嘧啶组,采用MTT法检测阿司匹林和氟尿嘧啶抑制结肠癌细胞增殖的效果,采用Hoechst 33258染色、caspase活性检测和流式细胞术检测研究药物诱发凋亡的作用及机制,采用real-time PCR和Western blotting方法检测药物对凋亡相关蛋白表达的调控作用。结果:氟尿嘧啶有效抑制HCT116和SW620结肠癌细胞增殖,且低浓度阿司匹林具有协同抑制的效果。氟尿嘧啶诱发HCT116细胞核出现明显的凋亡形态,caspase酶活性增高及sub-G1期比例增加。阿司匹林协同作用明显提高HCT116细胞凋亡的比例和caspase酶活性。此外,低浓度阿司匹林可以显著增强氟尿嘧啶抑制Bcl-2 mRNA及蛋白表达的效果。结论:阿司匹林和氟尿嘧啶具有协同抑制结肠癌细胞生长增殖的效果,且主要通过诱发细胞凋亡发挥作用。

AIM：To investigate the synergistic anti-proliferation effect of aspirin and 5-fluorouracil on the colon cancer cells and its mechanism .METHODS： Colon cancer cells were divided into 4 groups： control group , aspirin group, 5-fluorouracil group and aspirin ＋5-fluorouracil group .Synergistic anti-proliferation effect of aspirin and 5-fluorouracil on the colon cancer cells was observed by MTT assay .Apoptosis-inducing effect and mechanism were detected by Hoechst 33258 staining, caspase activity assay and flow cytometry analysis .The mRNA and protein levels of apoptosis-related proteins were evaluated by real-time PCR and Western blotting .RESULTS：5-Fluorouracil inhibited proliferation of HCT116 and SW620 colon cancer cells effectively , and low concentration of aspirin exerted synergistic inhibitory effect .5-Fluorouracil induced apoptotic morphology and increased caspase activity and sub -G1 phase in HCT116 cells.The synergistic effect of aspirin obviously enhanced apoptotic ratio and caspase activity .Moreover , 5-fluorouracil inhibited the mRNA and protein expression of Bcl-2, which was amplified by low concentration of aspirin .CONCLUSION：Aspirin and 5-fluorouracil had a synergistic anti-proliferation effect on the colon cancer cells through apoptosis pathway .