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基于基因表达谱芯片杂交分析Lm-PHB2转染前后CHL细胞的基因表达差异
时颖,郭思呈,李庆伟,李铁松
作者单位E-mail
时颖 辽宁师范大学生命科学学院 475640082@qq.com 
郭思呈 辽宁师范大学生命科学学院  
李庆伟 辽宁师范大学生命科学学院  
李铁松 辽宁师范大学生命科学学院  
摘要:
七鳃鳗是连接无脊椎动物与脊椎动物的重要桥梁,是脊椎动物进化生物学中重要的模式生物。为了检测抗增殖蛋白2(Prohibitin2,PHB2)基因进化水平,选取了10个物种与本课题组前期克隆得到的东北七鳃鳗PHB2(Lm-PHB2)进行氨基酸序列相似性对比,结果表明各物种的PHB2氨基酸序列在PHB结构域处高度保守,但在N-端和C-端氨基酸序列保守性较低。目前对人PHB2的研究较为广泛,但还未见Lm-PHB2转染人正常细胞后的基因表达差异的研究。因此利用基因芯片表达谱技术分析将重组质粒pEGFP-N1-Lm-PHB2瞬时转染入张氏肝(CHL)细胞后分析基因的表达差异。结果显示CHL细胞中共有270条显著差异表达基因,其中显著上调基因共141条,显著下调基因共129条,涉及细胞信号转导、细胞周期调节、细胞增殖、细胞代谢和细胞凋亡等多个方面。通过实时荧光定量PCR对基因表达谱芯片分析结果进行验证,结果显示转染pEGFP-N1-Lm-PHB2后细胞周期基因CDC25C、氧化应激相关基因(CAT、SOD、GST)和抗细胞凋亡基因HAX1均有显著性差异,与基因芯片表达谱结果一致。本研究首次应用基因表达谱芯片技术分析pEGFP-N1-Lm-PHB2转染到人细胞后基因的表达差异,这一研究不仅丰富了不同物种中PHB2的功能,而且为Lm-PHB2将来用来治疗人类乳腺癌、阿尔兹海默症和前列腺癌等疾病提供了充足且有价值的依据。
关键词:  七鳃鳗  PHB2  基因芯片表达谱  基因表达
DOI:
分类号:Q811.4
基金项目:国家自然科学(编号31501907)和辽宁省教育厅科学技术项目(编号:2015287)资助
Microarray analysis of gene expression profiling in CHL cells before and after Lm-PHB2 transfection
shiying guosicheng liqingwei litiesong,guosicheng,liqingwei,litiesong
Abstract:
Lamprey is an important bridge connecting invertebrates with vertebrates and important model organism in vertebrate evolutionary biology. In order to detect the evolutionary level of Prohibitin 2 (PHB2) gene,A total of 10 species were selected to compare the amino acid sequence similarity with the lamprey PHB2 (Lm-PHB2),which was obtained by cloning in our group. The results showed that the PHB2 amino acid sequences of the species were highly conserved at the PHB domain, but the sequence identity at the N-terminal and C-terminal amino acid sequences was low. At present, the amout of study about human PHB2 is more abundant, and the difference in gene expression after transfection of human normal cells with the recombinant plasmid pEGFP-N1-PHB2 has not been studied. Therefore, the difference of genes’ expression after the recombinant plasmid pEGFP-N1-PHB2 was transiently transfected into Chang liver cells(CHL)was detected by GeneChip Expression Profiling. The results showed that there were 270 significant differentially expressed genes in CHL cells, among them, a total of 141 genes were significantly up-regulated, and a total of 129 genes were down-regulated, including cell signal transduction, cell cycle regulation, cell proliferation, cell metabolism and apoptosis aspect. The results of gene chip expression were verified by real-time quantitative PCR. The results showed that after transfection of pEGFP-N1-Lm-PHB2, Cell cycle gene CDC25C, oxidative stress-related genes (CAT, SOD, GST) and anti-apoptotic gene HAX1 expression were significantly different. This result is consistent with the results of gene chip expression. In this study, the GeneChip Expression Profiling was used to analysis the the difference of genes’expression after transfection of Lm-PHB2 into human cells for the first time. This study not only enriched the function of PHB2 in different species, but also provide a sufficient and valuable basis for application of pEGFP-N1-Lm-PHB2 on the treatment of human breast cancer, Alzheimer's disease and prostate cancer and other diseases.
Key words:  Lampreys  PHB2  gene chip expression profiles  gene expression