XAF1基因诱导胰腺癌细胞株BxPC3凋亡的实验研究

目的 探讨Ad5/F35腺病毒介导的XAF1基因诱导胰腺癌细胞BxPC3凋亡及其可能的作用机制.方法 将前期构建的重组腺病毒Ad5/F35-XAF1感染胰腺癌细胞BxPc3.采用半定量RTPCR、蛋白质印迹检测法检测感染前后BxPC3细胞XAF1 mRNA和蛋白表达的变化;运用Annexin-V/PI和TUNEL法检测细胞的凋亡率;免疫蛋白印迹法检测细胞Caspase-3、PARP、Caspase-8和Bcl-2蛋白的表达.结果 Ad5/F35-XAF1感染后,BxPC3细胞的XAF1 mRNA和蛋白表达明显增高,与空白组和Ad5/F35-Null对照组比较,差异显著(P＜0.05);两种方法 检测的细胞凋亡率分别为(19.90±3.09)%、(9.29±2.13)%,较Ad5/F35-Null组的(6.72±0.76)%、(2.73±0.51)%和空白组的(7.22±1.53)%、(1.56±0.47)%均有显著差异(P＜0.01);Caspase-3、PARP、Caspase-8蛋白表达显著增强,Bcl-2表达降低.结论 Ad5F35-XAF1重组腺病毒感染胰腺癌细胞BxPC-3能明显诱导细胞的凋亡,其机制可能是激活死亡受体途径和线粒体途径.

XAF1 gene mediated by recombinant adenovirus induces apoptosis in pancreatic cell line BxPC-3

Objective To investigate whether expression of XAF1 mediated by edenovirus vector AdS/F35 could induce apoptosis in pancreatic cancer cell line BxPC-3 and its possible mechanisms.Methods Preconstructed recombinant Ad5/F35-XAF1 virus and negative control Ad5/F35-Null was tranfected into BxPC3:the expression of XAF1 mRNA and protein before and after tranfection was,analyzed by semi-quantitative RT-PCR and Western blot.The expressions of proteins including Caspase-3,PARP,Caspase-8 and bcl-2 were detected by Western blots.Cell apoptosis was assessed by Annexin-v/PI and TUNEL staining.Results After Ad5/F35-XAF1 tranfection,XAF1 mRNA and protein expression significantly increased,and the difference was statistically significant when compared with control group and Ad5/F35-Null group(P＜0.05).The apoptosis rate was(19.90±3.09)%and(9.29±2.13)%,which was significantly different(P＜0.01)from those in the Ad5/F35-Null group[(6.72±0.7)6%,(2.73±0.51)%]or in the control group[(7.22±1.53)%,(1.56±0.47)%].The expression of Caspase-3,PARP and Caspase-8 significantly increased,but the expression of bcl-2 decreased.Conclusions XAF1 plays a major role in the apoptosis of pancreatic cancer that acts thriugh the activation of death receptor pathway and mitochondrial pathway of apoptosis.