摘要
背景:课题组前期研究发现相较于野生型小鼠,线粒体蛋白磷酸酶2C(protein phosphatase 2Cm,PP2Cm)基因缺失小鼠明显发生肾功能衰竭的症状,因此推测PP2Cm可能在肾脏纤维化发展过程中发挥重要保护作用,然而其分子机制尚不明确。目的:探究PP2Cm基因对于人肾小管上皮细胞转录组的影响。方法:培养人肾小管上皮细胞,用质粒将PP2Cm基因转染进入人肾小管上皮细胞,荧光定量PCR实验和Western blot实验检测细胞中PP2Cm的表达,随后分别提取细胞RNA进行转录组测序,寻找转染组和对照组之间的差异性表达基因,利用生物信息学方法进一步对所得的差异基因进行GO分析和KEGG分析。结果与结论:通过测序分析,与未转染空白细胞相比,在转染PP2Cm基因的人肾小管上皮细胞中存在796个差异性表达基因,其中553个下调基因,243个上调基因,GO分析结果显示,上调表达的基因显著富集在细胞生物合成过程、蛋白质翻译、内在凋亡信号通路等;下调表达的基因显著富集在内皮细胞增殖、细胞黏附等信号通路;KEGG分析结果显示,显著上调表达的基因富集在氨基酸代谢、生物合成等代谢相关信号通路;下调表达的基因显著富集在泛酸和辅酶A的生物合成等信号通路。结果表明,PP2Cm过表达可以影响肾小管上皮细胞的一系列生物学过程相关的多条信号通路,可能在氨基酸代谢、生物合成等代谢相关信号通路中起重要作用。
BACKGROUND:A previous study by our group found that protein phosphatase 2Cm(PP2Cm)null mice developed significantly fewer symptoms of renal failure relative to wild-type mice,and thus it was speculated that PP2Cm may play an important protective role in the development of renal fibrosis,however,the molecular mechanisms remain undefined.OBJECTIVE:To investigate the effect of the PP2Cm gene on the transcriptome of human renal tubular epithelial cells.METHODS:Cultured human renal tubular epithelial cells were transfected with the PP2Cm gene into human renal tubular epithelial cells using plasmids.The expression of PP2Cm in the cells was detected by fluorescence quantitative PCR assay and western blot assay,and subsequently,cell RNA was separately extracted for transcriptome sequencing to look for differentially expressed genes between transfected and control groups.The resulting differential genes were further subjected to GO analysis and KEGG analysis using bioinformatics methods.RESULTS AND CONCLUSION:There were 796 differentially expressed genes,553 of which were downregulated genes and 243 upregulated genes,in human renal tubular epithelial cells transfected with the PP2Cm gene compared with untransfected blank cells by sequencing analysis.GO analysis results showed that the upregulated genes were significantly enriched in cellular biosynthetic processes,protein translation,intrinsic apoptotic signaling pathways,and so on.The downregulated expressed genes were significantly enriched in endothelial cell proliferation,cell adhesion and other signaling pathways.KEGG analysis results showed that the significantly up-regulated genes were enriched in metabolism-related signaling pathways such as amino acid metabolism and biosynthesis.The downregulated expressed genes were significantly enriched in signaling pathways such as pantothenate and coenzyme A biosynthesis.Our results show that PP2Cm overexpression can affect a number of signaling pathways related to a range of biological processes in renal tubular epithelial cells,which may be important in metabolism-related signaling pathways such as amino acid metabolism and biosynthesis.
作者
张丽
杨文君
桑晓红
韩媛媛
冒智捷
王顺
陆晨
Zhang Li;Yang Wenjun;Sang Xiaohong;Han Yuanyuan;Mao Zhijie;Wang Shun;Lu Chen(Nephrology Center of First Affiliated Hospital of Xinjiang Medical University,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Xinjiang Clinical Research Center of Renal Replacement Therapy,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Xinjiang Branch of National Clinical Research Center for Kidney Disease,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Xinjiang Blood Purification Medical Quality Control Center,Urumqi 830011,Xinjiang Uygur Autonomous Region,China;Institute of Nephrology of Xinjiang,Urumqi 830011,Xinjiang Uygur Autonomous Region,China)
出处
《中国组织工程研究》
CAS
北大核心
2024年第1期68-73,共6页
Chinese Journal of Tissue Engineering Research
基金
国家自然科学基金(81960145),项目负责人:张丽。
关键词
肾小管上皮细胞
PP2Cm
转录组测序
富集分析
信号通路
renal tubular epithelial cell
PP2Cm
transcriptome sequencing
enrichment analysis
signaling pathway
