基于生物信息学、分子对接与细胞实验探究C5orf46高表达在胃癌中的意义及潜在的中药干预

Significance of high expression of C5orf46 in gastric cancer and potential intervention of tarditional Chinese medicine based on bioinformatics,molecular docking,and cell experiments

该研究旨在探究C5orf46在胃癌中的表达、预后和临床意义,研究C5orf46的潜在有效成分与中药的相互作用。运用ggplot2包对胃癌组织和正常邻近组织中的C5orf46进行差异表达分析;运用survival包进行生存分析、单因素及多因素回归分析,运用Nomogram分析评估C5orf46在胃癌中的表达和总体生存率的关系;最后利用GSVA包对肿瘤浸润淋巴细胞丰度进行计算。Coremine数据库、TCMSP数据库与PubChem数据库用于查找C5orf46基因对应的潜在成分与中药成分,分子对接用于探讨潜在成分与C5orf46基因的结合强度,并利用细胞实验探究空白组、模型组、不同浓度给药组细胞C5orf46基因的相对表达情况。对比正常组织,胃癌组织中C5orf46的表达较高,在早期(T2期、N0期、M0期)有更显著的预测效应;肿瘤原发灶-区域淋巴结-远处转移(tumor-node-metastasis,TNM)分期越晚,C5orf46表达越高,胃癌患者生存概率越低;C5orf46的表达与胃癌中的辅助性T细胞1、巨噬细胞的浸润水平呈正相关趋势,与B细胞、中央记忆型T细胞、辅助性T细胞17、滤泡辅助性T细胞呈显著负相关趋势。C5orf46的对应潜在成分有7个,经过筛选得到3个有效成分,对应中药成分5个,分别为黑豆、大枣、瓜蒌、水飞蓟、天竺黄。分子对接显示唾液酸、单磷酸腺苷(adenosine monophosphate,AMP)与C5orf46结合情况较牢固。RTqPCR和Western blot结果显示,不同浓度AMP给药组C5orf46 mRNA和蛋白的表达水平均显著低于模型组,且浓度为40μmol·L^(-1)时表达水平最低。研究结果能为临床上治疗胃癌以及其他癌症的中药复方研发提供思路。

This study aims to investigate the expression,prognosis,and clinical significance of C5orf46 in gastric cancer and to study the interaction between the active components of C5orf46 and tarditional Chinese medicine.The ggplot2 package was utilized for differential expression analysis of C5orf46 in gastric cancer tissues and normal tissues.The survival package was used for survival analysis,univariate regression analysis,and multivariate regression analysis.Nomogram analysis was used to assess the connection between C5orf46 expression in gastric cancer and overall survival.The abundance of tumor-infiltrating lymphocytes was calculated by GSVA package.Coremine database,Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP)database,and PubChem database were used to search the potential components corresponding to C5orf46 gene and tarditional Chinese medicine.Molecular docking was performed to explore the binding affinity of potential components to C5orf46.Cell experiments were performed to explore the expression of C5orf46 gene in cells of the blank group,model group,and drug administration groups.As compared with normal tissues,C5orf46 expression was higher in gastric cancer tissues,which had more significant predictive effects in the early stages(T2,N0,and M0).The more advanced the tumor node metastasis(TNM)stage,the higher the C5orf46 expression and the lower the probability of survival of patients with gastric cancer.The expression of C5orf46 positively correlated with the helper T cells1 in gastric cancer and the macrophage infiltration level in gastric cancer,and negatively correlated with B cells,central memory T cells,helper T cells 17,and follicular helper T cells.Seven potential components of C5orf46 were obtained,and three active components were obtained after the screening,which matched five tarditional Chinese medicines,namely,Sojae Semen Nigrum,Jujubae Fructus,Trichosanthis Fructus,Silybi Fructus,and Bambusae Concretio Silicea.Molecular docking revealed that sialic acid and adenosine monophosphate(AMP)had a good binding ability to C5orf46.The results of real-time quantitative polymerase chain reaction(RT-qPCR)and Western blot showed that,as compared with the model group,the mRNA and protein expression levels of C5orf46 were significantly lower in the drug administration groups.The lowest expression level was found at the concentration of 40μmol·L^(-1).The results of this study provide ideas for the clinical development of traditional Chinese medicine compounds for the treatment of gastric cancer as well as other cancers.