Table_5_Transcriptomic and Proteomic Study on the High-Fat Diet Combined With AOM/DSS-Induced Adenomatous Polyps in Mice.xlsx

ObjectiveTo screen and identify molecular targets and bacteria genus leading to adenomatous polyps in mouse induced by high-fat diet (HFD) +AOM/DSS using omics technology.
MethodsThe molecular targets of colorectal adenoma disease were obtained from the GeneCards and OMIM database. The SPF C57BL mice were randomly divided into blank (Control) and AOM/DSS+HFD colorectal adenoma model (ADH) groups. The ADH model group was intraperitoneally injected with AOM reagent. Then, mice were given with 2.5% DSS (in free drinking water) and high-fat diet to establish the mouse model. During this period, the changes of physical signs of mice in each group were observed. After the end of modeling, HE staining was used to evaluate the histopathological change of mice. The differentially expressed genes and proteins in the Control group and ADH group were detected by RNA-seq transcriptome sequencing and Tandem Mass Tags (TMT) quantitative proteomics. The histological results were analyzed by intersection with the intestinal adenoma molecular targets obtained from the database. Moreover, the changes of intestinal flora in the two groups were examined. The correlation between targets and differential bacteria was analyzed and verified by Parallel Reaction Monitoring (PRM) to comprehensively evaluate the mouse model of adenomatous polyp induced by AOM/DSS+HFD.
ResultsThe general condition and histopathological results of mice confirmed that the ADH mouse model was successfully established and tubular adenoma was formed. A total of 604 genes and 42 proteins related to intestinal adenoma were obtained by histological analysis and database intersection analysis. The intestinal microflora of ADH mice was different from that of normal mice, and the constituents and abundance of intestinal flora were similar to those of human intestinal adenoma. GATA4 and LHPP were selected as potential pathological markers of the model mice by correlation analysis of targets and intestinal flora. The results of PRM verification were highly consistent with the results of RNA-Seq transcriptome sequencing and TMT analysis.
ConclusionThe pathological results, molecular pathological markers and the changes of intestinal flora suggest that the mouse ADH model is ideal for studying the transformation of inflammatory cancer. The ADH model will be helpful for understanding the occurrence and development of human colorectal cancer at the transcriptomic and proteomic level.

研究目的：本研究采用多组学技术，筛选并鉴定高脂饮食（high-fat diet, HFD）联合偶氮甲烷（azoxymethane, AOM）与葡聚糖硫酸钠（dextran sulfate sodium, DSS）诱导的小鼠腺瘤性息肉的潜在分子靶点与相关菌属。
研究方法：结直肠腺瘤疾病的分子靶点从GeneCards与OMIM数据库中获取。将无特定病原体（Specific Pathogen Free, SPF）级C57BL小鼠随机分为空白对照组（Control）与AOM/DSS+HFD结直肠腺瘤模型组（ADH）。模型组小鼠经腹腔注射AOM试剂，随后给予2.5% DSS（溶于自由饮水）与高脂饮食以构建小鼠模型。建模期间，每日观察各组小鼠的体征变化。建模结束后，采用苏木精-伊红（HE staining）染色评估小鼠的组织病理学变化。通过RNA-seq转录组测序（RNA-seq transcriptome sequencing）与串联质谱标签（Tandem Mass Tags, TMT）定量蛋白质组学技术，检测对照组与ADH组的差异表达基因与差异蛋白。将组织学结果与从数据库获取的肠腺瘤分子靶点进行交集分析，同时检测两组小鼠的肠道菌群变化。通过靶点与差异菌属的相关性分析，并采用平行反应监测（Parallel Reaction Monitoring, PRM）进行验证，以全面评估AOM/DSS+HFD诱导的腺瘤性息肉小鼠模型。
研究结果：小鼠的一般状态与组织病理学结果证实，ADH小鼠模型构建成功，且形成了管状腺瘤。通过组织学分析与数据库交集分析，共获得604个与肠腺瘤相关的基因与42个相关蛋白。ADH小鼠的肠道菌群与正常小鼠存在显著差异，其肠道菌群的组成与丰度模式与人类肠腺瘤相似。通过靶点与肠道菌群的相关性分析，筛选出GATA4与LHPP作为模型小鼠的潜在病理标志物。PRM验证结果与RNA-seq转录组测序及TMT定量蛋白质组学分析结果具有高度一致性。
研究结论：病理学结果、分子病理标志物及肠道菌群变化均表明，ADH小鼠模型是研究炎症致癌转化过程的理想模型。该ADH模型将有助于从转录组与蛋白质组层面解析人类结直肠癌的发生与发展机制。