摘要
目的探讨凋亡诱导因子(AIF)对人宫颈癌Hela细胞体外生物学活性的影响及其作用机制。方法通过脂质体转染pcDNA3空载体质粒及pcDNA3-AIF质粒于人宫颈癌Hela细胞,采用免疫荧光反应,Real time PCR及Western blot法检测细胞中AIF的表达。四甲基偶氮唑蓝(MTT)法检测细胞活力,Annexin V-PI流式双染及Hoechst染色检测细胞凋亡情况,Realtime PCR及Western blot法检测细胞中腺苷二磷酸核糖聚合酶-1(PARP-1)及核酸内切酶G(EndoG)蛋白及mRNA的表达。结果与pcDNA3组比较,pcDNA3-AIF组细胞中AIF主要定位于细胞核中,且细胞核中AIF蛋白及mRNA表达量皆显著提高(P<0.01),同时细胞活力下降,细胞凋亡率提高(P<0.01),细胞中PARP-1、EndoG蛋白及mRNA表达量显著提高(P<0.01)。结论 AIF具有促进宫颈癌Hela细胞凋亡的作用,可能与PARP-1/AIF/EndoG信号通路有关。
Objective To explore effect of apoptosis inducing factor（AIF）on the in vitro biological activity of human cervical cancer Hela cells and its mechanism. Methods Hela cells were transferred with pcDNA3 plasmid and peDNA3-AIF plasmid. The location and expression of AIF in Hela cell was determined by immunofluoreseent, realtime PCR and Western blot. The cell viability was detected by MTT. The celt apoptosis was examined by Annexin V-PI flow cytometry and Hoechst staining. The expression of t poly（ADP-ribose）polymerases-1（PARP-1）and endonuclease G（EndoG）was detected by Western blot. Results Compared with the pcDNA3 group, AIE in the pcDNA3-AIF group was mainly localizedin the nucleus, moreover the expression of AIF protein and mRNA in nueteus was significantly increased （P 〈0.01）, meanwhile the cell viability was decreased, cell apoptotic rate was in- creased（P〈0. 01） ,the expressions of PARP-1 and EndoG protein and mRNA were significantly increased（P〈0.01）. Conclusion AIF has.the effect for promoting Hela cell apoptosis,which might be related to P ARP-1/AIF/EndoG signal pathway.
出处
《重庆医学》
CAS
北大核心
2016年第32期4471-4474,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(81100394)
