Apaf-1基因转染及其对5-氟尿嘧啶诱导AGS细胞凋亡的影响

Apaf-1 gene transfer and its effects on apoptosis of AGS cell line

目的 Apaf 1是参与激活凋亡过程中Caspase系统的关键因子。研究Apaf 1在化疗药物诱导肿瘤细胞凋亡过程中的作用。方法 利用基因转染技术 ,将正、反义Apaf 1表达载体转染至AGS细胞系 ,建立Apaf 1过度表达及减低表达系统 ,观察在Apaf 1过度表达及减低表达情况下 ,5 氟尿嘧啶(5 FU)对AGS细胞凋亡过程的影响 ,并检测正、反义Apaf 1转基因AGS细胞株在 5 FU诱导的凋亡过程中细胞色素C信号转导通路相关分子的表达变化。结果 正义Apaf 1基因转染组cyt c、bax及cas pase 3基因的表达显著上调 ,而在反义Apaf 1基因转染组则相反。bcl 2基因的表达在正、反义Apaf 1基因转染组无明显变化。结论 过度表达Apaf 1基因可以明显增加AGS细胞对化疗药物 5 FU的敏感性 ,而Apaf

Objective Apaf-1 is a key factor involved in activating the Caspase system in apoptosis. The aim of this study was to evaluate the significance of Apaf-1 on the apoptosis process induced by chemotherapeutic agent. Methods We established Apaf-1 hyper- and hypo- expression system by transferred sense and anti-sense Apaf-1 cDNA into cultured AGS cell line. And meanwhile human cyt-c, Apaf-1, caspase-3, bax, and bcl-2 gene expressions were measured by RT-PCR method. Results Cyt-c, caspase-3, and bax gene expressions increased significantly after sense Apaf-1 transfection. On the contrary, anti-sense Apaf-1 transfer has minus effects on cyt-c, caspase-3, and bax gene expressions. No significant changes have been found on bcl-2 gene expression after transfection process.Conclusions Cyt-c pathway may play an important role in the apoptosis induced by chemotherapeutic drugs. The early expression of cyt-c and Apaf-1 may be the key procedure of cyt-c release from mitochondria and of caspases activation.