基于精准医学多维数据整合分析胃宁颗粒对胃癌甲基化网络的调控

It integrated analysis the regulation of gastric cancer methylation by Weining Granula base on base on precision medical multidimensional data ,The demethylation therapy will be developed rapidly in gastric cancer treatment, which due to methylation is an important kind of mechanism of epigenetic alterations in gastric cancer. The regulation of methylation by weining granula has been discovered in former NNSF of China (8116048). Weining granula inhibit the proliferation and induced the apoptosis of gastric carcinoma cell by intervening the process of methylation of molecule event in cell, which is a hypothesis proposed by the study. Accordingly, Screen whole-genome methylation site and its relational mRNA and protein as well as exploring what the weining granule have influence on gastric cancer methylation gene expression and transcription of mRNA base on using the SGC - 7910 cell model and precision medicine as well as law of biological DNA - mRNA - Protein center. Also according to Genome-transcriptome-proteome technology and methylation microarrays, expression profile chip as well as flight mass spectrum. Constructing the network model of methylation expression to enrich and cluster analysis by Cytoscape software in the light of interatomics. Analyze the complex treatment system and the interaction of biological response system by the research of methylation some-surface-stereo network. Explore the integrated mechanism of weining granula and its precise targets by its dynamic control process.

甲基化是胃癌发生发展的重要表观遗传学改变，去甲基化治疗将是胃癌治疗重要的发展方向。在前一国家自然科学基金（81160487）当中，我们发现胃宁颗粒对甲基化具有调控作用。该课题提出“胃宁颗粒通过干预甲基化过程细胞内分子事件，促进胃癌细胞凋亡，抑制胃癌细胞增殖”的科学假说,采用SGC-7910细胞模型，以精准医学理论为指导，按照生物DNA-mRNA -Proein中心法则，基于基因组-转录组-蛋白质组学技术，应用甲基化芯片、表达谱芯片、飞行质谱技术，筛查差异全基因组甲基化位点及其相关mRNA和蛋白，探索胃宁颗粒对胃癌甲基化基因表达、转录为mRNA，翻译成蛋白质的影响，根据相互作用组学，使用Cytoscape软件，构建甲基化表达调控网络模型，进行富集以及聚类分析，通过对甲基化点-面-立体网络的研究，分析复杂治疗系统和复杂生物应答系统的相互作用，以及动态调控过程，探索胃宁颗粒的整合机制和精准靶点。

本课题基于基因组-转录组-蛋白质组学技术，应用简化甲基化测序技术、定量蛋白质组学串联质谱标签技术及RNA-seq技术，筛查差异全基因组甲基化位点及其相关mRNA和蛋白，探索胃宁颗粒对胃癌甲基化基因表达、转录为mRNA，翻译成蛋白质的影响。从整体阐明胃宁颗粒对胃癌调控的整合机制和相关靶点。.基因组甲基化测序得到8983个差异甲基化区域（differentially methylated regions，DMRs），其中6234个高度甲基化区域，2749低甲基化区域。这些甲基化区域中，有1297个（14.44%）位于启动子，2109个（23.48%）位于内含子上，以及1029个（11.45%）位于外显子上。定量蛋白质组学串联质谱标签技术筛查差异蛋白，总定量蛋白6321个，显著差异表达蛋白191个，显著上调蛋白107个，显著下调蛋白84个。RNA-seq技术筛选差异表达基因，胃宁颗粒干预后，胃癌SGC-7901细胞株出现7902个显著差异表达基因，其中3886个（49.18%）基因表达上调，4016个（50.82%）基因表达下调。整合以上三大组学数据，利用String数据库构建PPI网络，并进行网络进行分析，鉴定出10个中枢基因:SOD2、HMOX1、MMP1、SRXN1、NOTCH1、MAPK14、TXNIP、VEGFA、POLR2F、HSPA9。这10个基因可能是胃宁颗粒影响胃癌甲基化基因表达、转录为mRNA，翻译成蛋白质的靶点基因。本课题从基因组学和蛋白质组学及转录组学角度阐明了胃宁颗粒的作用机制，为胃宁颗粒干预胃癌的提供了科学依据。