NKX3.1下调前列腺癌PC-3细胞中抗凋亡基因bcl-2的表达  

NKX3.1 down-regulates bcl-2 gene expression in prostate cancer PC-3 cells

作　　者：陈兆波[1] 刘春艳[1] 倪娜娜[1] 于洋[1] 张鹏举[1] 陈蔚文[1] 崔福爱[1] 姜安丽[1] 

机构地区：[1]山东大学医学院生物化学与分子生物学研究所,山东济南250012

出　　处：《中国病理生理杂志》2011年第10期1902-1906,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81071720);山东省优秀中青年科学家科研奖励基金计划资助项目(No.BS2009SW042);山东省科技发展计划资助项目(No.2009GG20002045)n

摘　　要：目的:研究前列腺特异转录因子NKX3.1对bcl-2基因表达以及前列腺癌细胞凋亡的影响。方法:转染pcDNA3.1-NKX3.1于前列腺癌PC-3细胞中,得到瞬时转染NKX3.1的PC-3细胞,通过RT-PCR和Western blotting方法研究NKX3.1对bcl-2基因的表达的影响,并通过EMSA技术,研究NKX3.1诱导bcl-2基因下调的分子机制;进一步通过流式细胞术、凋亡小体染色等方法研究NKX3.1对细胞凋亡的影响。结果:NKX3.1可明显下调PC-3细胞中bcl-2 mRNA及Bcl-2蛋白的表达;EMSA结果证明NKX3.1可与bcl-2基因上游调控区中的NKX3.1结合元件相互作用;流式细胞术结果显示,PC-3细胞转染NKX3.1后,细胞凋亡数增加了1.41倍;用Hoechst 33258细胞染色发现NKX3.1可使PC-3细胞凋亡小体数量明显增加。结论:NKX3.1可下调抗凋亡基因bcl-2的表达,促进前列腺癌细胞凋亡。AIM：To investigate the effect of NKX3.1 on the gene expression of bcl-2 and apoptosis in prostate cancer PC-3 cells.METHODS： The eukaryotic expression plasmid pcDNA3.1-NKX3.1 was transiently transfected into PC-3 cells.RT-PCR and Western blotting were used to detect the effects of NKX3.1 on the expression of bcl-2 gene.Down-regulatory effect of NKX3.1 on bcl-2 gene was detected by EMSA.Flow cytometry and apoptotic body staining were carried out to study the effects of NKX3.1 on apoptosis of PC-3 cells.RESULTS： The mRNA and protein expression of bcl-2 in PC-3 cells was down-regulated by over-expression of NKX3.1.The EMSA result showed that NKX3.1 interacted with the NKX3.1 binding elements in upstream regulatory region of bcl-2 gene.The results of flow cytometry showed that the number of apoptotic PC-3 cells increased by 1.41-fold after NKX3.1 transfection to PC-3 cells.NKX3.1 increased the apoptotic bodies stained by Hoechst 33258 significantly.CONCLUSION： NKX3.1 down-regulates the expression of anti-apoptotic gene bcl-2 and induces the apoptosis of prostate cancer PC-3 cells.

关 键 词：NKX3.1蛋白 PC-3细胞 基因 BCL-2 细胞凋亡 

分 类 号：R737.25[医药卫生—肿瘤]