Mao Jiajie, Xu Chen, Hu Dongsheng, Bao Huijing
Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
The 4th Central Hospital, Laboratory Science Department, Tianjin, 300100, China.
Naunyn Schmiedebergs Arch Pharmacol. 2025 Apr 28. doi: 10.1007/s00210-025-04200-0.
Cinobufotalin (CINO), a bioactive compound derived from toad skin secretions, has demonstrated clinical efficacy in cancer treatment. However, its molecular mechanisms in ovarian cancer (OC) remain poorly characterized. This study systematically investigated the anti-OC mechanisms of CINO through an integrated strategy combining network pharmacology, molecular docking, and RNA sequencing. Potential CINO targets were predicted via Swiss Target Prediction, while OC-related genes were retrieved from GeneCards and OMIM. Intersecting targets were analyzed using PPI networks and functional enrichment (GO/KEGG). Molecular docking validated ligand-target interactions, and RNA-seq quantified differential gene expression in CINO-treated SKOV3 cells. Among 69 overlapping targets, 10 hub genes (EGFR, PTGS2, MDM2, MAPK1, MAPK3, MTOR, ESR1, PIK3CA, MMP9, and GSK3B) were identified. KEGG analysis highlighted the MAPK signaling and endocrine resistance pathways. RNA-seq revealed 1488 upregulated and 3253 downregulated DEGs, which were mainly enriched in axon development, axonogenesis, and primarily involved in the MAPK signaling pathway. CINO significantly suppressed EGFR, ESR1, MAPK1, MDM2, and mTOR expression (p < 0.05), aligning with pathway predictions. CINO exerts anti-OC effects by modulating endocrine resistance and MAPK signaling, providing a mechanistic foundation for its clinical application.
华蟾酥毒基（CINO）是一种从蟾蜍皮肤分泌物中提取的生物活性化合物，已在癌症治疗中显示出临床疗效。然而，其在卵巢癌（OC）中的分子机制仍不清楚。本研究通过整合网络药理学、分子对接和RNA测序的策略，系统地研究了CINO的抗OC机制。通过瑞士靶点预测工具预测潜在的CINO靶点，同时从基因卡片（GeneCards）和在线人类孟德尔遗传数据库（OMIM）中检索OC相关基因。使用蛋白质-蛋白质相互作用（PPI）网络和功能富集分析（基因本体论/京都基因与基因组百科全书，GO/KEGG）对交集靶点进行分析。分子对接验证配体-靶点相互作用，RNA测序定量CINO处理的SKOV3细胞中的差异基因表达。在69个重叠靶点中，鉴定出10个核心基因（表皮生长因子受体，EGFR；前列腺素内过氧化物合酶2，PTGS2；小鼠双微体2，MDM2；丝裂原活化蛋白激酶1，MAPK1；丝裂原活化蛋白激酶3，MAPK3；哺乳动物雷帕霉素靶蛋白，MTOR；雌激素受体1，ESR1；磷脂酰肌醇-3激酶催化亚基α，PIK3CA；基质金属蛋白酶9，MMP9；糖原合成酶激酶3β，GSK3B）。KEGG分析突出了丝裂原活化蛋白激酶信号通路和内分泌抵抗途径。RNA测序显示1488个基因上调和3253个基因下调，这些基因主要富集在轴突发育、轴突形成中，并主要参与丝裂原活化蛋白激酶信号通路。CINO显著抑制EGFR、ESR1、MAPK1、MDM2和MTOR的表达（p<0.05），与通路预测结果一致。CINO通过调节内分泌抵抗和丝裂原活化蛋白激酶信号通路发挥抗OC作用，为其临床应用提供了机制基础。
