ERK_(1,2)抑制剂联合5-FU对B16细胞增殖和凋亡的影响

Effects of the combination of the MEK_(1,2) inhibitor and 5-FU on proliferation and apoptosis of B16 cells

目的:观察ERK1,2抑制剂联合5-FU对B16细胞增殖和凋亡的影响,并探讨作用机制。方法:用MTT法观察ERK1,2抑制剂、5-FU和联合用药对细胞增殖的抑制作用,流式细胞仪检测细胞凋亡率,用RT-PCR观察ERK1,2抑制剂、5-FU和联合用药对bcl-2和caspase-9的表达的影响。结果:ERK1,2抑制剂联合5-FU组在抑制细胞增殖,诱导细胞凋亡,均较对照组、单独用药组作用增强,并且下调bcl-2和上调caspase-9的表达。结论:ERK1,2抑制剂联合5-FU有协同抑制B16细胞增殖,促进凋亡的作用,其机制可能与诱导细胞凋亡,下调bcl-2和上调caspase-9的表达有关。

Aim：To study the effects of combination of the ERK1,2 inhibitor and 5-FU on B16 cells in vitro and their antitumor mechanisms. Methods： To detect the growth inhibitory effects of the specific inhibitor of MAPK/ERK1,2, 5-FU alone or combination in various concentration on B16 cells by MTY assys. Flow cytometry was used to determine the effects of the specific inhibitor of MAPK/ERK1,2, 5-FU alone or combination in various concentration on apoptosis was ananlyzed by using flow cytometry. The reverse transcription polymerase chain reaction was used to detect the influence of the specific inhibitor of MAPK/ERK1,2, 5-FU alone or combination to the expression of Bcl-2 and Caspase-9. Results： Combination of the specific inhibitor of MAPK/ERK1.2 withS-FU had greater inhibitory effects on growth and proliferation, enhanced the effects of cellular apoptosis of B16 cells, and decreased the expression of bcl-2 and increased the expression of caspase-9, compared to the sum of the specific inhibitor of MAPK/ ERK1,2, 5-FU alone and the control group. Conclusion： Combination of specific inhibitor of MAPK/ ERK1,2 with5-FU has a significantly synergic anti cancer effect. The antitumor mechanism may be associated with induction of apoptosis and down-regulation of anti-apoptotic bcl-2 and up-regulation of the caspase-9.