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中国癌症防治杂志

中国癌症防治杂志 ›› 2023, Vol. 15 ›› Issue (1): 18-24.doi: 10.3969/j.issn.1674?5671.2023.01.03

• 基础研究 • 上一篇    下一篇

扁塑藤素对膀胱癌细胞增殖和凋亡的影响及其作用机制

  

  1. 遵义医科大学附属医院；成都大学附属医院泌尿外科
  • 出版日期:2023-02-25 发布日期:2023-03-01
  • 通讯作者: 陈林 E-mail： z15928562618@163.com
  • 基金资助:
    国家自然科学基金项目（81500577）；四川省科学技术厅科研课题（2022NSFSC1592）；成都市卫生健康委员会医学科研课题（YP201501）；贵州省优秀青年科技人才培养对象专项基金项目［黔科合人字（2015）31号］；成都优秀卫技人才发展专项资金资助项目；成都大学附属医院攀登人才计划项目（RCJH?01?04）

Effects of pristimerin on proliferation and apoptosis of bladder cancer cells and its mechanism

  • Online:2023-02-25 Published:2023-03-01

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摘要： 目的 探讨扁塑藤素对人膀胱癌HT⁃1376细胞增殖和凋亡的影响及其可能的作用机制。 方法 通过TCMSP、SwissTargetPrediction、PharmMapper数据库共同预测扁塑藤素的作用靶点，从GeneCards数据库获取膀胱癌靶点；通过Venny 2.1.0 平台整合扁塑藤素治疗膀胱癌的潜在靶点，利用String平台、Cytoscape 3.9.1软件构建蛋白质⁃蛋白质相互作用（protein⁃protein interaction，PPI）网络，通过DAVID 数据库进行GO功能和KEGG通路富集分析。体外培养人正常膀胱上皮细胞SV⁃HUC⁃1和人膀胱癌细胞株HT⁃1376，用0~0.8 μmol/L扁塑藤素处理后，采用CCK⁃8实验、菌落形成实验以及Hoechst⁃PI染色实验检测扁塑藤素对HT⁃1376细胞增殖及凋亡的影响，采用Western blot检测凋亡相关蛋白Bcl⁃2、Bax、Cleaved PARP、PARP以及MEK/ERK信号通路相关蛋白p⁃MEK、p⁃ERK1/2的表达水平。结果 扁塑藤素的作用靶点有292个，其中与膀胱癌治疗相关的潜在靶点有114个，这些靶点共涉及434种生物过程、54种细胞组分、114种分子功能，主要富集于MAPK信号通路、脂质与动脉粥样硬化等132条信号通路。体外实验验证结果显示，扁塑藤素能显著抑制HT⁃1376细胞的活力和集落形成能力，并诱导细胞凋亡（均P<0.05）。Western blot实验显示，与未加药处理的对照组相比，0.65 μmol/L扁塑藤素处理后HT⁃1376细胞中凋亡相关蛋白Bax和Cleaved PARP的表达水平升高，Bcl⁃2和PARP的表达水平降低，MEK/ERK信号通路相关蛋白p⁃MEK、p⁃ERK1/2表达水平也明显降低（均P<0.05）。 结论 扁塑藤素可能通过调控MEK/ERK信号通路相关蛋白的磷酸化水平抑制膀胱癌细胞增殖并诱导细胞凋亡。

关键词: 膀胱癌, 扁塑藤素, 网络药理学, 增殖, 凋亡, MEK/ERK

Abstract: Objective To investigate the effects of pristimerin on the proliferation and apoptosis of human bladder cancer HT⁃1376 cells and the related mechanisms. Methods The target protein of pristimerin was predicted by TCMSP, SwissTargetPrediction database and PharmMapper database, and the target of bladder cancer was obtained from GeneCards database. The potential targets of pristimerin in the treatment of bladder cancer were integrated through the Venny 2.1.0 platform; the String database and Cytoscape 3.9.1 software were used to construct a protein⁃protein interaction (PPI) network. GO function and KEGG pathway enrichment analyses were performed by DAVID database. The normal human bladder epithelial cells SV⁃HUC⁃1 and human bladder cancer cell line HT⁃1376 cells were cultured in vitro, and treated with 0-0.8 μmol/L pristimerin. The effects of pristimerin on the proliferation and apoptosis of HT⁃1376 cells were detected by CCK⁃8 assay, colony formation assay, Hoechst⁃PI staining assay. Western blot was used to detect the expression levels of apoptosis⁃related proteins Bcl⁃2, Bax, Cleaved PARP, PARP, and MEK/ERK signaling pathway⁃related proteins p⁃MEK, p⁃ERK1/2. Results  There were 292 targets of pristimerin, among which 114 potential targets were related to bladder cancer treatment, involving 434 biological processes, 54 cellular components and 114 molecular functions, mainly enriched in 132 signaling pathways such as MAPK signaling pathway, lipid and atherosclerosis, etc. The results of in vitro experimental verification showed that pristimerin could significantly inhibit the viability and colony formation ability of HT⁃1376 cells and induce cell apoptosis(all P<0.05). Western blot showed that compared with untreated control group, the expression levels of apoptosis⁃related proteins Bax and Cleaved PARP were increased, Bcl⁃2 and PARP were decreased, and MEK/ERK signaling pathway⁃related proteins p⁃MEK and p⁃ERK1/2 were decreased after treated with 0.65 μmol/L pristimerin (all P<0.05). Conclusions Pristimerin may inhibit the proliferation and induce apoptosis of bladder cancer cells by regulating the phosphorylation level of MEK/ERK signaling pathway⁃related proteins.

Key words: Bladder Cancer, Pristimerin, Network pharmacology, Proliferation, Apoptosis, MEK/ERK

中图分类号: 

  • R737.14