转录组与蛋白组关联分析探讨草酸钙结石晶体对HK-2细胞基因表达的影响  

Proteotranscriptomic analyses of calcium oxalate monohydrate-stimulated HK-2 cells

作　　者：张圣平[1] 邓琼[1] 张颖[1] 张建文[1] 梁辉[1] 王铸 ZHANG Shengping;DENG Qiong;ZHANG Ying;ZHANG Jianwen;LIANG Hui;WANG Zhu(Department of Urology,Affiliated Hospital of Longhua Shenzhen,Southern Medical University,Shenzhen 518109,China)

机构地区：[1]南方医科大学附属深圳龙华医院泌尿外科,广东深圳518109

出　　处：《现代泌尿外科杂志》2022年第11期927-931,共5页Journal of Modern Urology

基　　金：深圳市科技计划项目(No.JCYJ20210324131414039);深圳市龙华区科技创新项目(No.LHKJCXJCYJ202002);深圳市龙华区医疗卫生机构科研项目(No.2020003、No.2020013)。

摘　　要：目的探讨草酸钙结石晶体对人近曲肾小管上皮细胞HK-2的影响。方法建立草酸钙结石与HK-2细胞相互作用的模型,进行转录组和蛋白组表达谱高通量定量研究。筛选差异表达基因,通过InterProScan进行GO富集分析,KEGG Mapper进行KEEG信号通路分析及蛋白质相互作用网络STRING分析。结果转录组与蛋白组定量研究分别定量到57833个转录本和6407个蛋白。通过比较转录组与蛋白组的定量情况,发现有6144个基因在转录组和蛋白组水平都定量。蛋白水平464个分子显著下调,699个分子显著上调。转录组水平11个分子下调,14个分子上调。GO分析发现,线粒体衍生囊泡与腺嘌呤跨膜转运体活性、snRNA的细胞核输出以及中心粒周围异染色质组装具有最高的富集指数。KEEG富集分析显示缬氨酸、亮氨酸和异亮氨酸降解、初级胆汁酸生物合成、维生素B6代谢、硫中继系统以及糖胺聚糖生物合成和DNA非同源末端链接为最显著的KEEG通路。蛋白互作网络发现MYH9、MYH14、MYH1、MYL12A、ACTB、ARPC1A、ARPC2、ANXA2、CFL1、PRDX1、PRXD2共11个关键调控因子。结论转录组与蛋白组关联分析筛选所得关键通路及关键基因可能在肾结石形成过程中扮演重要角色。Objective To investigate the effects of calcium oxalate stone crystal on human renal proximal tubular epithe-lial HK-2 cells.Methods The calcium oxalate crystal-HK-2 cell interaction model was established,and characterized by trans-criptome and proteome analysis.The differentially expressed genes were screened and analyzed by bioinformatic methods.InterProScan and KEGG Mapper were used to perform GO and KEGG pathway enrichment analysis.The protein interaction network(PPI)was constructed by STRING.Results A total of 57833 transcripts and 6407 proteins were quantified in trans-criptome and proteome quantitative studies,respectively;6144 genes were quantified at both transcriptome and proteome levels;464 molecules were significantly down-regulated and 699 molecules were significantly up-regulated at proteome level.At the transcriptome level,11 molecules were down-regulated and 14 molecules were up-regulated.Mitochondrion-derived vesicle,adenine transmembrane transporter activity,snRNA export from nucleus and pericentric heterochromatin assembly showed the highest GO enrichment factor.Valine,leucine and isoleucine degradation and primary bile acid biosynthesis,vitamin B6 meta-bolism and sulfur relay system were the most enriched KEEG pathways.Altogether 11 key genes were identified by SRING analysis,including MYH9,MYH14,MYH1,MYL12 A,ACTB,ARPC1 A,ARPC2,ANXA2,CFL1,PRDX1 and PRDX2.Conclusion The key genes and pathways screened by proteotranscriptomic analysis may play important roles in the formation of renal calculi.

关 键 词：草酸钙结石 转录组 蛋白组 近曲肾小管上皮细胞 肾结石 

分 类 号：R692.4[医药卫生—泌尿科学]