基于加权基因共表达网络分析和分子对接分析荔枝核干预结肠腺癌的黄酮类成分及靶点

目的 探讨荔枝核干预结肠腺癌（COAD）进展和转移的黄酮类成分和作用靶点。方法 通过DRAR-CPI和SWISS数据库检索荔枝核中19个黄酮类化合物的潜在作用靶点。从TCGA数据库下载COAD基因表达数据和临床特征数据，采用加权基因共表达网络分析（WGCNA）法建立COAD的基因共表达网络和识别共表达模块，以共表达模块和潜在作用靶点中共同靶点作为化合物干预COAD的作用靶点。通过String数据库进行蛋白互作网络分析、KEGG和GO分析。通过cytoHubba插件提取Hub基因作为COAD潜在生物标志物，通过Cytoscape建立成分、靶点、通路的互作网络。HPA数据库验证潜在生物标志物的表达，通过分子对接技术虚拟筛选与潜在生物标志物作用的化合物。结果 经WGCNA分析得到18个共表达模块，其中7个模块与生存时间、肿瘤分期等临床特征相关，turquoise模块与COAD进展转移相关。荔枝核中19个黄酮类化合物作用于380个潜在作用靶点，选择与turquoise模块重复的34个靶点作为作用靶点，GO分析结果显示作用靶点富在304个GO条目，其中生物过程229条、细胞组成31条、分子功能44条；KEGG分析结果显示作用靶点富集在癌症通路、细胞周期、孕酮介导的卵母癌症途径、细胞衰老、p53信号通路等40条通路。CytoHubba筛选得到CDC25A、CDC25C、CCNB2、AURKB基因作为COAD进展转移相关的潜在生物标志物。HPA数据库免疫组化结果显示，与癌旁组织相比，COAD组织中CDC25C、AURKB、CCNB2蛋白表达升高（P<0.05），与TCGA数据集中基因表达量一致。通过分子对接初步筛选得到柚皮芸香苷、原花青素A2、根皮苷、表儿茶素可通过氢键等与CDC25A、CDC25C、AURKB结合较好。结论 CDC25A、CDC25C、CCNB2、AURKB可作为COAD进展和转移密切相关的潜在生物标志物。荔枝核黄酮类化合物干预COAD进展和转移的机制可能与黄酮类化合物调控细胞分裂、细胞周期G2/M期转变等生物过程进而调节癌症途径、p53信号通路等信号通路有关，其中柚皮芸香苷、原花青素A2、根皮苷、表儿茶素、芦丁可能是CDC25A、CDC25C、AURKB的潜在抑制剂。

Screening of flavonoids in Litchi Semen and its targets of intervention in colonic adenocarcinoma by weighted gene co-expression network analysis and molecular docking

Objective To screen the flavonoid constituents and targets of Litchi Semen in the intervention of progression and metastasis of colon adenocarcinoma (COAD). Methods Through DRAR-CPI and SWISS database, potential targets of 19 flavonoids in Litchi Semen were searched. COAD gene expression data and clinical characteristic data from TCGA database were downloaded. Weighted gene co-expression network analysis (WGCNA) was used to establish the gene co-expression network and identify the co-expression module of COAD. The common targets of co-expression module and potential targets were used as the compound to interfere with the drug target of COAD. Protein interaction network analysis, KEGG and GO analysis were performed by String database. The Hub gene was extracted as potential biomarkers of COAD by the cytoHubba, and the interaction network of components, targets and pathways was established by the Cytoscape. The expressions of potential biomarkers were verified by HPA database, and the compounds were docked with the potential biomarkers. Results A total of 18 co-expression modules were identified with seven of them were correlated with clinical features, such as survival time and tumor stage. Turquoise module was related to the development and transfer of COAD. 19 flavonoids in Litchi Semen acted on 380 potential targets. 34 targets repeated with turquoise module were selected as targets. GO analysis showed that the target points were enriched in 304 GO items, including 229 biological processes, 31 cell composition and 44 molecular functions; KEGG analysis showed that target points were enriched in cancer pathways, cell cycle, and progesterone-mediated 40 pathways including oocyte cancer pathway, cell senescence, and p53 signaling pathway. The genes of CDC25A, CDC25C, CCNB2 and AURKB were screened by cytoHubba as potential biomarkers which related to the progress and transfer of COAD. Compared with para-cancerous tissues, immunohistochemistry results obtained from HPA database showed that the protein expressions of CDC25C, AURKB and CCNB2 in COAD were increased significantly (P<0.05), which were consistent with gene expression in TCGA data set. Narirutin, procyanidin A2, phloridzin and ent-epicatechin which were well combined to CDC25A, CDC25C and AURKB through hydrogen bond were screened. Conclusion CDC25A, CDC25C, CCNB2 and AURKB were the potential biomarkers closely related to the progression and metastasis of COAD. The mechanism of intervention of flavonoids in Litchi Semen on the progression and metastasis of COAD may be related to the regulation of biological processes, such as cell division, G2/M phase transformation of cell cycle, and the regulation of cancer pathway, p53 signaling pathway and other signaling pathways. Narirutin, procyanidin A2, phloridzin, ent-epicatechin and rutin could be treated as potential inhibitors of CDC25A, CDC25C and AURKB.