Hermawan Adam, Hanif Naufa, Putri Dyaningtyas Dewi Pamungkas, Fatimah Nurul, Prasetio Heri Himawan
Laboratory of Macromolecular Engineering, Department of Pharmaceutical Chemistry, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Laboratory of Advanced Pharmaceutical Sciences, Faculty of Pharmacy, Universitas Gadjah Mada, Yogyakarta, Indonesia.
Discov Oncol. 2025 Mar 20;16(1):365. doi: 10.1007/s12672-025-02116-y.
Breast cancer is a potentially fatal illness that affects millions of women worldwide. Methotrexate (MTX) may be beneficial for treating breast cancer; however, high doses and prolonged use can cause drug resistance. Although certain citrus flavonoids-nobiletin, sinensetin, tangeretin, hesperidin, hesperetin, and naringenin-may overcome resistance to chemotherapy, no study has investigated MTX resistance. This study investigated the potential of natural chemicals, specifically nobiletin and sinensetin, to overcome MTX resistance in breast cancer cells using MTX-resistant MCF-7 (MCF-7/MTX) and MCF-7 cells. Protein targets of citrus flavonoids were identified from multiple databases and were collected using Venny 2.1. Microarray data of MCF-7 and MCF-7/MTX cells were acquired from the Gene Expression Omnibus. Subsequently, we constructed a protein-protein interaction network and selected the hub proteins. Gene ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis, drug- and disease-gene enrichment analyses, genetic alteration examination, receiver operating characteristic curve analysis, mRNA levels analysis, prognostic value analysis, and molecular docking analysis were performed along with in vitro experiments. Cytotoxicity of citrus flavonoids (individually and combined) was assessed in MCF-7/MTX cells. Nobiletin and sinensetin significantly enhanced the cytotoxicity of MTX in MCF-7/MTX cells. BCL2L1, CDK1, EGFR, PTGS2, PLK1, MMP2, ACHE, ABCG2, and KIT genes were enriched in cholinesterase activity, cell cycle regulation, and the PI3K/Akt signaling pathway. Nobiletin and sinensetin impeded PLK1, CDK1, and ACHE activities based on molecular docking. Nobiletin and sinensetin in combination with MTX may overcome breast cancer cell resistance to MTX.
乳腺癌是一种潜在的致命疾病，影响着全球数百万女性。甲氨蝶呤（MTX）可能对治疗乳腺癌有益；然而，高剂量和长期使用会导致耐药性。尽管某些柑橘类黄酮——川陈皮素、甜橙黄酮、橘红素、橙皮苷、橙皮素和柚皮素——可能克服化疗耐药性，但尚无研究调查MTX耐药性。本研究使用耐MTX的MCF-7（MCF-7/MTX）细胞和MCF-7细胞，研究天然化学物质，特别是川陈皮素和甜橙黄酮，克服乳腺癌细胞中MTX耐药性的潜力。从多个数据库中鉴定柑橘类黄酮的蛋白质靶点，并使用Venny 2.1进行收集。MCF-7和MCF-7/MTX细胞的微阵列数据从基因表达综合数据库中获取。随后，我们构建了蛋白质-蛋白质相互作用网络并选择了枢纽蛋白。进行了基因本体论和京都基因与基因组百科全书通路富集分析、药物和疾病基因富集分析、基因改变检查、受试者工作特征曲线分析、mRNA水平分析、预后价值分析和分子对接分析以及体外实验。评估了柑橘类黄酮（单独和联合使用）对MCF-7/MTX细胞的细胞毒性。川陈皮素和甜橙黄酮显著增强了MTX对MCF-7/MTX细胞的细胞毒性。BCL2L1、CDK1、EGFR、PTGS2、PLK1、MMP2、ACHE、ABCG2和KIT基因在胆碱酯酶活性、细胞周期调控和PI3K/Akt信号通路中富集。基于分子对接，川陈皮素和甜橙黄酮可抑制PLK1、CDK1和ACHE的活性。川陈皮素和甜橙黄酮与MTX联合使用可能克服乳腺癌细胞对MTX的耐药性。
