ARHI对5-FU诱导胃癌细胞周期阻滞和凋亡的影响

摘要

Objective To elucidate the effects of ARHI on 5-FU-induced gastric cancer cell cycle arrest and apoptosis. Methods BGC-823 gastric cancer cells were stably transfected with recombinant plasmid vector pIRES2-EGFP or pIRES2-EGFP-ARHI for the ex-periments. After 5-fluorouracil(5-FU) stimulation, flow cytometry was used to detect the changes of cell cycle and apoptosis. Western blot was used to detect the protein expression levels of cyclin D1 ,Bax,Bcl-2 and activated caspase-3. Results Both ARHI and 5-FU induced G1 phase arrest in BGC-823 cells,and the percentage of G1 cells increased to 61. 64%± 4. 56%(P<0. 05) and 65. 71%± 4. 89%(P<0. 01), respectively. The combined effect of ARHI and 5-FU was more significant, and the percentage of G1 cells in-creased to 81. 14%± 5. 63%(P<0. 01). ARHI and 5-FU-induced G1 phase arrest was accompanied by down-regulated protein expres-sion of cyclin D1. ARHI had no effect on BGC-823 cell apoptosis,and 5-FU was able to induce significant apoptosis with an apoptotic rate of 7. 22%± 1. 55%(P<0. 01). The combined effect of ARHI and 5-FU was more significant, and the apoptotic rate increased to 17. 34%± 2. 81%(P<0. 01). 5-FU alone or ARHI and 5-FU-induced BGC-823 cell apoptosis were accompanied by increased Bax/Bcl-2 ratio and activated caspase-3 expression. Conclusion ARHI can enhance 5-FU-induced cell cycle arrest and apoptosis in gas-tric cancer cells.%目的：探讨ARHI对5-FU诱导胃癌细胞周期阻滞和凋亡的影响。方法以稳定转染pIRES2-EGFP或pIRES2-EG-FP-ARHI重组质粒的BGC-823胃癌细胞系为模型,采用5-氟尿嘧啶(5-FU)刺激细胞24 h,通过流式细胞术检测细胞周期和凋亡的改变,利用Western blot检测cyclin D1、Bax、Bcl-2及活化caspase-3蛋白的表达水平。结果 ARHI与5-FU都能够导致BGC-823细胞发生G1期阻滞,G1期比例分别增加至61.64%±4.56%(P<0.05)和65.71%±4.89%(P<0.01),二者联合作用效果更明显,G1期比例增加至81.14%±5.63%(P<0.01)；ARHI与5-FU引起的G1期阻滞与cyclin D1的表达水平降低相关；ARHI对BGC-823细胞凋亡没有影响,5-FU可导致细胞凋亡,凋亡率为7.22%±1.55%(P<0.01),而二者联合作用导致更显著的细胞凋亡,凋亡率达到17.34%±2.81%(P<0.01)；5-FU单独或与ARHI联合促使Bax/Bcl-2比值增高、caspase-3的活化增强。结论 ARHI能够增强5-FU诱导的胃癌细胞周期阻滞和凋亡。