pokemon通过抑制p53和上调白细胞介素6抑制胃肿瘤细胞BGC823凋亡  

Pokemon inhibits the apoptosis of gastric cancer cell line BGC823 by suppressing p53 and enhancing IL-6

作　　者：季玉连[1] 赵树立[1] 赵光峰[1] 侯亚义[1,2] 

机构地区：[1]南京大学医学院免疫与生殖生物实验室 [2]南京大学医药生物技术国家重点实验室

出　　处：《医学研究生学报》2012年第11期1155-1159,共5页Journal of Medical Postgraduates

摘　　要：目的原癌基因pokemon是转录抑制因子,高表达于多种肿瘤细胞中,但其在肿瘤发生发展中的机制仍有待阐明。文中研究pokemon在胃腺癌细胞系BGC823中的作用及机制。方法将pokemon的编码片段构建入真核表达载体pcD-NA3.1(+)中。将具有靶向沉默pokemon的siRNA互补的DNA构建到pSilencer 3.1 H1-neo中。将构建的质粒pcDNA3.1-pokemon和pSilencer 3.1 H1-3#用脂质体2000转染BGC823细胞。用流式细胞仪检测细胞凋亡,用RT-PCR分析细胞内mRNA的变化,用Western blot检测pokemon蛋白的表达。结果成功构建了pokemon过表达/干扰载体pcDNA3.1-pokemon和pSi-lencer 3.1 H1-3#。在BGC823细胞中,过表达的pokemon可显著抑制p53,促进白细胞介素6(IL-6)转录。干扰pokemon基因的表达可使p53的转录增强。pokemon过表达可抑制BGC823的凋亡,沉默pokemon表达可促进凋亡。结论 pokemon抑制胃肿瘤细胞BGC823凋亡,p53和IL-6是pokemon下游的靶基因。Objective The proto-oncogene Pokemon is a potent transcriptional repressor highly expressed in many kinds of cancers,but its role in the development and progression of cancer remains to be clarified.In this study,we investigated the role of Pokemon in gastric cancer cell line BGC823 and its mechanism.Methods The coding sequence of Pokemon was constructed in the eukaryotic expression vector pcDNA3.1（＋）,and a specific siRNA targeting Pokemon was constructed in pSilencer 3.1 H1-neo.The resultant recombinant plasmids pcDNA3.1-pokemon and pSilencer 3.1 H1-#3 were transfected into the BGC823 cells using Lipofectamine 2000.The apoptosis of the cells was evaluated by flow cytometry,and the mRNA and protein expressions of Pokemon detemined by PCR analysis and Western blot,respectively.Results We successfully constructed the gene overexpression and interruption vectors pcDNA3.1-pokemon and pSilencer 3.1 H1-#3.Overexpressed Pokemon significantly inhibited p53 and enhanced IL-6,while the interference of the Pokemon expression resulted in an increased transcription of p53.The overexpression of Pokemon suppressed,while knockdown of Pokemon increased the apoptosis of BGC823 cells.Conclusion Pokemon inhibits the apoptosis of gastric cancer cell line BGC823.p53 and IL-6 are the downstream target genes of Pokemon.

关 键 词：POKEMON P53 IL-6 细胞凋亡 BGC823细胞 

分 类 号：R735.2[医药卫生—肿瘤]