白藜芦醇联合伊立替康治疗结直肠癌的分子作用机制研究

Molecular mechanism of resveratrol combined with irinotecan in treatment of colorectal cancer

该研究旨在探讨白藜芦醇(resveratrol,RES)联合伊立替康(irinotecan,IRI)治疗结直肠癌(colorectal cancer,CRC)的作用机制。笔者借助数据库分别获取RES、IRI及CRC的靶点,作韦恩图得到RES联合IRI治疗CRC的作用靶点,并对其进行蛋白功能聚类、GO和KEGG富集分析。构建蛋白互作(PPI)网络图,并进行核心靶基因筛选与靶点-信号通路网络图构建;借助IGEMDOCK对核心靶基因分子对接。分析关键靶基因表达水平与CRC预后、免疫浸润的关系,并利用体外细胞实验探究分析RES联合IRI治疗CRC的相关分子机制。结果显示共获得63个RES联合IRI治疗CRC的潜在靶点,且蛋白功能聚类分析发现约23%为跨膜信号受体、22%为蛋白质修饰酶、14%靶点为代谢物相互转化酶。这些靶基因GO生物过程(BP)主要富集于蛋白质自身磷酸化,GO细胞组分(CC)主要富集于受体复合物和细胞膜等,GO分子功能(MF)主要富集于跨膜受体蛋白酪氨酸激酶活性;KEGG信号通路主要富集于中心碳代谢。RES联合IRI治疗CRC的关键靶点为PIK3CA、EGFR、IGF1R,这三者均与CRC免疫浸润显著正相关,分子对接结果显示PIK3CA与RES、IRI的结合最稳定。相比于对照组,RES单药处理组、IRI单药处理组及RES+IRI联合处理组CRC细胞的增殖能力和EGFR蛋白表达水平均显著降低;且RES+IRI联合处理组CRC细胞的细胞增殖能力和EGFR蛋白表达水平均显著低于IRI单药处理组。综上,PIK3CA、EGFR、IGF1R是RES联合IRI治疗CRC的关键作用靶点,RES可通过下调EGFR信号通路转导抑制结直肠癌细胞增殖并改善其IRI化疗耐药性。

This study aimed to investigate the mechanism of resveratrol(RES)combined with irinotecan(IRI)in the treatment of colorectal cancer(CRC).The targets of RES,IRI,and CRC were obtained from databases,and the targets of RES combined with IRI in the treatment of CRC were acquired by Venn diagram.The protein functional cluster analysis,GO and KEGG enrichment analyses were performed.In addition,the protein-protein interaction(PPI)network was constructed.The core target genes were screened out and the target-signaling pathway network was set up.IGEMDOCK was used to dock the core target gene molecules.Besides,the relationship between the expression level of key target genes and the prognosis and immune infiltration of CRC was analyzed.Based on the in vitro cell experiment,the molecular mechanism of RES combined with IRI in the treatment of CRC was explored and analyzed.According to the results,63 potential targets of RES combined with IRI were obtained for CRC treatment.Furthermore,cluster analysis revealed that protein functions included 23%transmembrane signal receptors,22%protein modifying enzymes,and 14%metabolite converting enzymes.GO analysis indicated that BPs were mainly concentrated in protein autophosphorylation,CCs in receptor complex and plasma membrane,and MFs in transmembrane receptor protein tyrosine kinase activity.Moreover,KEGG signaling pathways were mainly enriched in central carbon metabolism in cancer.The key targets of RES combined with IRI in the treatment of CRC were PIK3CA,EGFR,and IGF1R,all of which were significantly positively correlated with the immune infiltration of CRC.As shown by the molecular docking results,PIK3CA had the most stable binding with RES and IRI.Compared with the results in the control group,the proliferation ability and EGFR protein expression of CRC cells in the RES-treated group,the IRI-treated group,and the RES+IRI treated group significantly decreased.Moreover,the cell proliferation ability and EGFR protein expression level of CRC cells in the RES+IRI treated group were remarkably lower than those in the IRI-treated group.In conclusion,PIK3CA,EGFR,and IGF1R are the key targets of RES combined with IRI in CRC treatment.In addition,RES can inhibit the proliferation of CRC cells and improve IRI chemoresistance by downregulating the EGFR signaling pathway.