通过网络药理学与生物信息学及实验验证相结合的方法来阐明榅桲总多酚(total polyphenols of Cydonia oblonga Mil-ler，TPCOM)抗肾癌的作用机制。通过网络药理学筛选榅桲活性多酚类化合物和肾癌相关的靶点。通过对Gene Expression Omnibus(GEO)数据库中肾癌患者肿瘤组织和正常组织RNA测序数据进行差异表达基因分析，结合网络药理学预测结果和差异表达基因分析获得TPCOM作用于肾癌的核心基因，结合生存分析找出能够影响患者生存的关键靶点，进行京都基因与基因组百科全书(Kyoto Encyclopedia of Genes and Genomes，KEGG)和基因本体(Gene Ontology，GO)富集分析。采用 20～640μg·mL-1 TPCOM 处理 786-O 和 Renca 细胞进行 cell counting kit-8(CCK-8)实验，40、80、160 μg·mL-1 TPCOM 处理肾癌细胞，检测TPCOM对肾癌细胞迁移和肾癌细胞中蛋白激酶B(protein kinase B，Akt)、雷帕霉素靶蛋白(mammalian target of rapamycin，mTOR)和磷脂酰肌醇3-激酶(phosphoinositide 3-kinase，PI3K)信号通路相关蛋白表达水平的调控作用。通过网络药理学预测获得8个榅桲多酚类活性化合物，通过生存分析获得TPCOM发挥抗肾癌作用的15个显著影响患者生存的肾癌差异表达基因。KEGG和GO分析结果显示以上15个靶点主要与PI3K/Akt信号通路、细胞迁移和增殖等生物过程有关。结果显示TP-COM 能够抑制786-O和Renca细胞的增殖，IC50分别为121。4、137。9 μg·mL-1。TPCOM能够抑制786-O和Renca细胞的迁移，能够抑制PI3K/Akt/mTOR信号通路。榅桲总多酚可能通过抑制PI3K/Akt/mTOR信号通路的激活从而抑制肾癌细胞的增殖和迁移来发挥抗肾癌作用。该研究为新疆天然产物榅桲的抗肿瘤作用研究提供一定的研究基础，对进一步推动新疆榅桲的开发和利用具有拓展和补充意义。
Total polyphenols of Cydonia oblonga inhibited proliferation and migration of renal cancer cells by PI3K/Akt/mTOR pathway
The mechanism of total polyphenols of Cydonia oblonga Miller(TPCOM)against kidney cancer was elucidated through a combination of network pharmacology,bioinformatics,and experimental verification.The active polyphenolic compounds from C.ob-longa were screened by network pharmacological techniques and kidney cancer-related targets were collected through the database.The differential gene expression analysis was performed on RNA sequencing data from tumor tissue and normal tissue of kidney cancer pa-tients obtained from the Gene Expression Omnibus(GEO)database.The results of network pharmacology predictions and differential gene expression analysis were used to identify the core genes targeted by TPCOM in kidney cancer.Survival analysis was conducted to identify key targets that could impact patient survival,followed by Kyoto Encyclopedia of Genes and Genomes(KEGG)and Gene On-tology(GO)enrichment analyses.Cell proliferation and activity experiments(cell counting kit-8)were conducted using TPCOM at concentrations ranging from 20 to 640 μg·mL-1 on 786-O and Renca cells.Additionally,TPCOM at concentrations of 40,80,and 160μg·mL-1 was applied to kidney cancer cells to assess its effect on cell migration and its regulation of protein expression levels related to the protein kinase B(Akt),mammalian target of rapamycin(mTOR),and phosphoinositide 3-kinase(PI3K)signaling pathways.Network pharmacology predicted eight active polyphenolic compounds from C.oblonga.Survival analysis revealed 15 significantly dif-ferentially expressed genes in kidney cancer that were affected by TPCOM and had a significant impact on patient survival.KEGG and GO analysis results indicated that these 15 targets were primarily associated with the PI3K/Akt signaling pathway,cell migration,and proliferation.The results showed that TPCOM could inhibit the proliferation of 786-O and Renca cells,with IC50 values of 121.4 and 137.9 μg·mL-1,respectively.TPCOM was also found to inhibit the migration of these cells and suppress the PI3K/Akt/mTOR signa-ling pathway.TPCOM may exert its anti-kidney cancer effects by inhibiting the activation of the PI3K/Akt/mTOR signaling pathway,thereby restraining the proliferation and migration of kidney cancer cells.This study provides a foundation for the research on the anti-tumor effects of natural product C.oblonga,particularly in Xinjiang,and holds significance for further promoting its development and utilization.
total polyphenols of Cydonia oblongakidney cancerbioinformaticsnetwork pharmacology
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