基于iTRAQ技术分析LCMT1高表达对人胚肾细胞的影响

Effect of Overexpression of LCMT1 on Human Embryonic Kidney Cells by iTRAQ and tandem mass spectrometry

LCMT1是重要的哺乳动物甲基转移酶,为了探索LCMT1在生物体内可能涉及的生物进程,并建立LCMT1高表达的差异表达蛋白谱。我们成功构建了LCMT1高表达和质粒对照细胞株,利用同位素标记相对和绝对定量（isobaric tags for relative and absolute quantitation,i TRAQ）技术鉴定LCMT1高表达引起的差异表达蛋白,并进行了生物信息学分析。我们共鉴定到4 480个蛋白,差异表达蛋白233个,其中上调蛋白78个,下调蛋白155个。涉及到的细胞生物进程5条,信号通路15条。蛋白质相互作用分析结果表明差异表达蛋白主要分为4群,这些差异表达蛋白主要与RNA翻译与泛素化,肾脏发育,羧酸代谢过程相关。本研究发现LCMT1高表达引起了大量蛋白的差异表达,而这些差异表达蛋白的识别为后续LCMT1相关疾病的研究提供了丰富的理论依据。

LCMT1 is an important mammal methyltransferase.To preliminarily explore the possible biological process related to LCMT1 in organism and establish the differentially expression protein spectrums of overexpression of LCMT1.We successful constructed LCMT1 overexpression and vector control cells by plasmid transfection technique.Differentially expression proteins were identified by isobaric tags for relative and absolute quantitation（i TRAQ）.Finally,bioinformatics analysis of the data was carried out.Totally,4 480 proteins were identified,including 233 differentially expressed proteins（78 proteins were upregulated and 155 proteins were downregulated）.These proteins were involved in 5 biological process,15 signaling pathway.Analysis of protein-protein interaction indicated that differentially expression proteins were mainly divided into 4 groups.These differentially expression proteins were probably involved in translational,ubiquitination,kidney development,carboxylic acid metabolic process.In this study,we found that Overexpression of LCMT1 induced many differentially expression proteins,and these proteins were identified provides a theoretical evidence for the study of LCMT1-related diseases.