首页|基于网络药理学和体外实验验证木犀草素抑制膀胱癌细胞的增殖

基于网络药理学和体外实验验证木犀草素抑制膀胱癌细胞的增殖

Verification of the Inhibition Effect of Luteolin on the Proliferation of Bladder Cancer Cells Based on Network Pharmacoloev and Cell Experiments

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  • 国家科技期刊平台
  • NETL
  • NSTL
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目的:用网络药理学和体外实验验证的方法探讨木犀草素抑制膀胱癌细胞增殖的作用机制.方法:使用中药系统药理学数据库与分析平台、SwissTarget Prediction数据库、Pharmmapper数据库和GeneCards数据库,筛选出木犀草素和膀胱癌的作用靶点,利用Venny2.1取出交集靶点,用交集靶点进行基因本体(GO)和京都基因与基因组百科全书(KEGG)分析并利用String数据库构建蛋白质-蛋白质相互作用网络,最后用Cytoscape构建成分-通路-靶点-疾病网络图.通过细胞计数试剂盒-8(CCK8)、集落形成实验检测膀胱癌细胞的活力以及集落形成能力、蛋白质免疫印迹检测促分裂原活化的蛋白激酶(MAPK)通路上蛋白的表达以及增殖细胞核抗原(PCNA)的表达,5-乙炔基-2'-脱氧尿苷(EDU)实验进一步检测木犀草素对膀胱癌细胞增殖的影响.结果:通过网络药理学一共筛出136个木犀草素与膀胱癌的潜在靶点,GO分析中与生物过程相关主要涉及491个功能簇,与细胞组分相关主要涉及52个功能簇,与分子功能相关主要涉及112个功能簇.KEGG通路富集分析共获得151条通路,主要有癌症通路、PI3K-AKT信号通路、MAPK信号通路.体外实验表明木犀草素通过MAPK信号通路有效抑制了膀胱癌细胞HT1376的增殖;CCK8实验结果表明,木犀草素能明显抑制膀胱癌细胞HT1376的活力;集落形成实验表明木犀草素具有明显抑制膀胱癌细胞集落形成的能力;蛋白质免疫印迹实验结果表明,木犀草素能明显抑制MAPK通路上P-RAF、P-MEK、P-ERK1/2蛋白的表达,并且降低了 PCNA蛋白的表达水平;EDU实验表明木犀草素能明显抑制膀胱癌HT1376细胞的增殖.结论:本实验通过网络药理学和体外实验验证的方法证明了木犀草素可能通过MAPK信号通路抑制膀胱癌HT1376细胞的增殖.
Objective:To evaluate the mechanism of luteolin in inhibiting the proliferation of bladder cancer cells by means of net-work pharmacology and cell experiments.Methods:The targets of luteolin and bladder cancer were retrieved from the Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP),SwissTarget Prediction,Pharmmapper,and GeneCards,and Venny2.1 was used to extract the common targets shared by luteolin and bladder cancer.The Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analyses were performed for the common targets,and the pro-tein-protein interaction network was constructed via String.Cytoscape was used to build an active compound-pathway-target-disease network.The viability and colony formation ability of bladder cancer cells were examined by the Cell Counting Kit-8(CCK8)and colony formation assays,respectively.The expression levels of the proteins in the mitogen-activated protein kinase(MAPK)signa-ling pathway and proliferating cell nuclear antigen(PCNA)were determined by Western blotting.The 5-ethynyl-2'-deoxyuridine(EDU)assay was employed to detect the effect of luteolin on the proliferation of bladder cancer cells.Results:A total of 136 po-tential targets of luteolin and bladder cancer were screened out by network pharmacology.The GO analysis predicted 491,52,and 112 functional clusters involved in biological processes,cellular components,and molecular functions,respectively.A total of 151 pathways were obtained by the KEGG pathway enrichment analysis,including cancer pathway,phosphatidylinositol 3-kinase(P13K)-protein kinase B(AKT)signaling pathway,and MAPK signaling pathway.The results of cell experiments showed that lute-olin inhibited the proliferation of bladder cancer HT1376 cells through the MAPK signaling pathway.The results of the CCK8 assay showed that luteolin inhibited the viability of HT1376 cells.The results of the colony formation assay showed that luteolin inhibited the ability of bladder cancer cells to form colonies.Western blotting showed that luteolin significantly inhibited the expression of phospho-rapidly accelerated fibrosarcoma(P-RAF),phospho-mitogen-activated extracellular signal-regulated kinase(P-MEK),and phospho-extracellular regulated kinase 1/2(p-ERK1/2)in the MAPK pathway and down-regulated the expression of PCNA.The EDU assay showed that luteolin significantly inhibited the proliferation of HT1376 cells.Conclusion:Luteolin may inhibit the pro-liferation of bladder cancer HT1376 cells via the MAPK signaling pathway,as indicated by network pharmacology and cell experi-ments.

LuteolinMitogen-activated protein kinase(MAPK)signaling pathwayBladder cancerCell proliferationNetwork pharmacologyHT1376 cells

代文斌、陈林、周鑫、陈波、王威威、龙晓明、董陶涛、梁国标、杨进

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遵义医科大学,遵义,563000

成都大学附属医院泌尿外科,成都,610081

木犀草素 促分裂原活化的蛋白激酶信号通路 膀胱癌 细胞增殖 网络药理学 HT1376细胞

国家自然科学基金项目成都市卫生健康委员会医学科研课题贵州省优秀青年科技人才培养对象专项基金成都优秀卫技人才发展专项成都大学附属医院攀登人才计划项目

81500577YP201501黔科合人字[2015]31号RCJH-01-04

2024

10.3969/j.issn.1673-7202.2024.10.002

世界中医药
世界中医药学会联合会

世界中医药

CSTPCDCHSSCD北大核心
影响因子：1.266
ISSN：1673-7202
年,卷(期)：2024.19(10)
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