PDF(Pubmed)
Abstract：
BACKGROUND: Cepharanthine, a bioactive constituent of Stephania japonica (Thunb.) Miers, is known for its potent anti-tumor properties. Nevertheless, the precise impact of this substance on bladder cancer remains poorly comprehended. The aim of this study was to demonstrate the effect and mechanism of cepharanthine on the metastasis of human bladder cancer cells.
METHODS: The application of network pharmacology was utilized to ascertain the possible targets and signaling pathways of cepharanthine in the treatment of bladder cancer. The antiproliferative effects of cepharanthine were evaluated using Cell Counting Kit-8 and colony formation assays. The migration and invasion capabilities were assessed using Transwell assays and wound healing experiments. Proteins related to the Rap1 signaling pathway, cellular migration, cellular invasion, and Epithelial-Mesenchymal Transition (EMT) were quantified by western blotting.
RESULTS: Through database screening, 313 cepharanthine-acting targets, 277 candidate disease targets in bladder cancer, 22 intersecting targets, and 12 core targets were confirmed. The involvement of the Rap1 signaling system was revealed by the Kyoto Encyclopedia of Genes and Genomes\' pathway enrichment study. Cepharanthine was shown to decrease bladder cancer cell proliferation, migration, and invasion in vitro. Cepharanthine activated the Rap1 signaling pathway by upregulating Epac1 and downregulating E-cadherin and C3G protein expression, leading to increased expression of Rap1 GTP protein and decreased expression of protein kinase D1 and integrin α5. Rap1 signalling pathway activation resulted in the downregulation of migration and invasion-related proteins, matrix metallopeptidase MMP2, MMP9, as well as EMT-related proteins, N-cadherin and Snail, without affecting vimentin expression.
CONCLUSIONS: Cepharanthine inhibits migration, invasion, and EMT of bladder cancer cells by activating the Rap1 signalling pathway. The results offer helpful insights regarding the possible therapeutic use of cepharanthine for treating bladder cancer.
METHODS: The application of network pharmacology was utilized to ascertain the possible targets and signaling pathways of cepharanthine in the treatment of bladder cancer. The antiproliferative effects of cepharanthine were evaluated using Cell Counting Kit-8 and colony formation assays. The migration and invasion capabilities were assessed using Transwell assays and wound healing experiments. Proteins related to the Rap1 signaling pathway, cellular migration, cellular invasion, and Epithelial-Mesenchymal Transition (EMT) were quantified by western blotting.
RESULTS: Through database screening, 313 cepharanthine-acting targets, 277 candidate disease targets in bladder cancer, 22 intersecting targets, and 12 core targets were confirmed. The involvement of the Rap1 signaling system was revealed by the Kyoto Encyclopedia of Genes and Genomes\' pathway enrichment study. Cepharanthine was shown to decrease bladder cancer cell proliferation, migration, and invasion in vitro. Cepharanthine activated the Rap1 signaling pathway by upregulating Epac1 and downregulating E-cadherin and C3G protein expression, leading to increased expression of Rap1 GTP protein and decreased expression of protein kinase D1 and integrin α5. Rap1 signalling pathway activation resulted in the downregulation of migration and invasion-related proteins, matrix metallopeptidase MMP2, MMP9, as well as EMT-related proteins, N-cadherin and Snail, without affecting vimentin expression.
CONCLUSIONS: Cepharanthine inhibits migration, invasion, and EMT of bladder cancer cells by activating the Rap1 signalling pathway. The results offer helpful insights regarding the possible therapeutic use of cepharanthine for treating bladder cancer.
摘要：
背景：西法兰碱，Stephaniajaponica（Thunb。)Miers,以其有效的抗肿瘤特性而闻名。然而，这种物质对膀胱癌的确切影响仍然知之甚少。本研究的目的是证明西黄嘌呤对人膀胱癌细胞转移的影响和机制。
方法：利用网络药理学的应用来确定西黄嘌呤治疗膀胱癌的可能靶点和信号通路。使用细胞计数试剂盒-8和集落形成测定来评价西黄嘌呤的抗增殖作用。使用Transwell测定和伤口愈合实验评估迁移和侵袭能力。与Rap1信号通路相关的蛋白质，细胞迁移，细胞入侵，和上皮-间充质转化(EMT)通过蛋白质印迹定量。
结果：通过数据库筛选，313头孢甘辛作用靶点，277个膀胱癌候选疾病靶点，22个相交目标，并确认了12个核心目标。《京都基因和基因组百科全书》途径富集研究揭示了Rap1信号系统的参与。研究表明，黄芩苷可降低膀胱癌细胞的增殖，迁移,和体外侵袭。通过上调Epac1并下调E-cadherin和C3G蛋白表达激活Rap1信号通路,导致Rap1GTP蛋白表达增加，蛋白激酶D1和整合素α5表达降低。Rap1信号通路激活导致迁移和侵袭相关蛋白下调,基质金属肽酶MMP2，MMP9，以及EMT相关蛋白，N-钙黏着蛋白和蜗牛，不影响波形蛋白表达。
结论：西法兰碱抑制迁移，入侵,和EMT的膀胱癌细胞通过激活Rap1信号通路。该结果提供了有关西法兰碱治疗膀胱癌的可能治疗用途的有用见解。
方法：利用网络药理学的应用来确定西黄嘌呤治疗膀胱癌的可能靶点和信号通路。使用细胞计数试剂盒-8和集落形成测定来评价西黄嘌呤的抗增殖作用。使用Transwell测定和伤口愈合实验评估迁移和侵袭能力。与Rap1信号通路相关的蛋白质，细胞迁移，细胞入侵，和上皮-间充质转化(EMT)通过蛋白质印迹定量。
结果：通过数据库筛选，313头孢甘辛作用靶点，277个膀胱癌候选疾病靶点，22个相交目标，并确认了12个核心目标。《京都基因和基因组百科全书》途径富集研究揭示了Rap1信号系统的参与。研究表明，黄芩苷可降低膀胱癌细胞的增殖，迁移,和体外侵袭。通过上调Epac1并下调E-cadherin和C3G蛋白表达激活Rap1信号通路,导致Rap1GTP蛋白表达增加，蛋白激酶D1和整合素α5表达降低。Rap1信号通路激活导致迁移和侵袭相关蛋白下调,基质金属肽酶MMP2，MMP9，以及EMT相关蛋白，N-钙黏着蛋白和蜗牛，不影响波形蛋白表达。
结论：西法兰碱抑制迁移，入侵,和EMT的膀胱癌细胞通过激活Rap1信号通路。该结果提供了有关西法兰碱治疗膀胱癌的可能治疗用途的有用见解。
