摘要
目的:探讨含有AFP启动子的携蜂毒素基因重组腺病毒诱导HepG2细胞凋亡的相关机制。方法:采用AnnexinV/PI双染法,流式细胞仪检测含rAFP的蜂毒素基因重组腺病毒(Ad-rAFP-Mel)感染HepG2细胞后的凋亡情况;采用罗丹明123、PI双染色法检测线粒体膜电位的变化,Fura-2染色检测细胞内Ca2+变化。结果:Ad-rAFP-Mel感染HepG2细胞后发生明显的细胞凋亡,线粒体膜电位下降,细胞内钙离子的浓度升高。结论:结果提示感染后表达的内源性蜂毒素可能通过影响线粒体、内质网等结构,从而诱导AFP阳性的肝癌细胞凋亡。
Objective：To investigate the effects of recombinant adenovirus harboring melittin gene,under the control of transcription regulatory element of alpha-fetoprotein（Ad-rAFP-Mel）,on apoptosis rate of hepatocarcinoma cell line HepG2 and its related mechanism.Methods：Cell apoptosis was confirmed by detecting phosphatidylserine（PS）externalization with Annexin V/PI double staining.Rhodamine 123 and PI dyes were used to check the changes of mitochondrial membrane potential of the Ad-rAFP-Mel infected HCC cells determined by FCM,while Fura-2 dye was used to detect the intracellular calcium ion concentration determined by FCM too,both of them demonstrated the state of biomenbrane.Results：Ad-rAFP-Mel promoted cell apoptosis of hepatocarcinoma cell line HepG2 in a dose-dependent and time-dependent manner.Ad-rAFP-Mel could lead to the reducing of mitochondrial membrane potential and the increasing of intracellular calcium ion concentration.Conclusion：Ad-rAFP-Mel can induce cell apoptosis,this phenomena may be explained through the changes of biomembrane structure caused by it.
出处
《中西医结合肝病杂志》
CAS
2010年第5期283-286,共4页
Chinese Journal of Integrated Traditional and Western Medicine on Liver Diseases
基金
国家自然科学基金资助项目(No.30600808
No.30772887)
