简介：由于当前治疗方法的局限性及其相关的不良副作用，宫颈癌是全球女性死亡的主要原因之一。 Launaea cornuta 被用作治疗包括癌症在内的多种疾病的传统药物。然而，目前还没有科学验证角藻对宫颈癌的抗增殖活性。目的：本研究旨在通过网络分析方法评估角花草的选择性抗增殖、细胞毒和抗迁移作用，并探讨其对人宫颈癌细胞系（HeLa-229）的治疗机制。材料和方法：通过 3-(4, 5-二甲基噻唑-2-基)-2, 5-二苯基溴化四唑 (MTT) 生物测定法和通过伤口愈合测定评估抗迁移作用。使用定性颜色法和气相色谱-质谱法 (GC-MS) 分析化合物。随后，通过生物信息学分析，包括蛋白质-蛋白质相互作用（PPI）网络分析、基因本体论（GO）和京都基因与基因组百科全书（KEGG）分析，筛选羊角草潜在的抗宫颈癌治疗靶基因。 。进行分子对接（MD）是为了预测和了解配体对抗宫颈癌的分子相互作用。进行逆转录定量聚合酶链反应（RT-qPCR）来验证网络分析结果。结果：L. cornuta 乙酸乙酯组分对 HeLa-229 增殖表现出显着的细胞毒性作用（IC50 为 20.56 ± 2.83 µg/mL），选择性指数 (SI) 为 2.36，对非癌细胞的细胞毒性最小 (Vero-CCL 81) （IC50 为 48.83 ± 23.02）。通过 GC-MS 分析，还鉴定出 124 个与 L.获得了cornuta乙酸乙酯组分对人宫颈癌的影响，其中AKT1、MDM2、CDK2、MCL1和MTOR被确定为顶级枢纽基因，PI3K/Akt1、Ras/MAPK、FoxO和EGFR信号通路被确定为显着富集的通路。分子对接结果显示，豆甾基甲基醚对CDK2、ATK1、BCL2、MDM2和Casp9具有良好的结合亲和力，结合能范围为-7.0至-12.6 kcal/mol Tremulone与TP53和P21具有良好的结合亲和力。分别为-7.0 和-8.0 kcal/mol。这表明角藻化合物的乙酸乙酯部分与选定的宫颈癌靶基因存在稳定的分子相互作用。此外，RT-qPCR 分析显示，与阴性对照（DMSO 0.2%）相比，用 L. cornuta 处理的 HeLa-229 细胞中 CDK2、MDM2 和 BCL2 下调，Casp9 和 P21 上调。结论：研究结果表明，角花草醋酸乙酯部分植物化学物质可调节各种分子靶点和途径，对 HeLa-229 细胞表现出选择性抗增殖和细胞毒性作用。这项研究为进一步研究开发创新的临床抗宫颈癌药物奠定了基础。版权所有 © 2024 Lagu、Nyamai 和 Njeru。

Introduction: Cervical cancer is one of the leading causes of death among women globally due to the limitation of current treatment methods and their associated adverse side effects. Launaea cornuta is used as traditional medicine for the treatment of a variety of diseases including cancer. However, there is no scientific validation on the antiproliferative activity of L. cornuta against cervical cancer. Objective: This study aimed to evaluate the selective antiproliferative, cytotoxic and antimigratory effects of L. cornuta and to explore its therapeutical mechanisms in human cervical cancer cell lines (HeLa-229) through a network analysis approach. Materials and methods: The cytotoxic effect of L. cornuta ethyl acetate fraction on the proliferation of cervical cancer cells was evaluated by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) bioassay and the antimigratory effect was assessed by wound healing assays. Compounds were analysed using the qualitative colour method and gas chromatography-mass spectroscopy (GC-MS). Subsequently, bioinformatic analyses, including the protein-protein interaction (PPI) network analysis, Gene Ontology (GO), and Kyoto Encyclopaedia of Genes and Genomes (KEGG) analysis, were performed to screen for potential anticervical cancer therapeutic target genes of L. cornuta. Molecular docking (MD) was performed to predict and understand the molecular interactions of the ligands against cervical cancer. Reverse transcription-quantitative polymerase chain reaction (RT-qPCR) was performed to validate the network analysis results. Results: L. cornuta ethyl acetate fraction exhibited a remarkable cytotoxic effect on HeLa-229 proliferation (IC50 of 20.56 ± 2.83 μg/mL) with a selectivity index (SI) of 2.36 with minimal cytotoxicity on non-cancerous cells (Vero-CCL 81 (IC50 of 48.83 ± 23.02). The preliminary screening revealed the presence of glycosides, phenols, saponins, terpenoids, quinones, and tannins. Thirteen compounds were also identified by GC-MS analysis. 124 potential target genes associated with the effect of L. cornuta ethyl acetate fraction on human cervical cancer were obtained, including AKT1, MDM2, CDK2, MCL1 and MTOR were identified among the top hub genes and PI3K/Akt1, Ras/MAPK, FoxO and EGFR signalling pathways were identified as the significantly enriched pathways. Molecular docking results showed that stigmasteryl methyl ether had a good binding affinity against CDK2, ATK1, BCL2, MDM2, and Casp9, with binding energy ranging from -7.0 to -12.6 kcal/mol. Tremulone showed a good binding affinity against TP53 and P21 with -7.0 and -8.0 kcal/mol, respectively. This suggests a stable molecular interaction of the ethyl acetate fraction of L. cornuta compounds with the selected target genes for cervical cancer. Furthermore, RT-qPCR analysis revealed that CDK2, MDM2 and BCL2 were downregulated, and Casp9 and P21 were upregulated in HeLa-229 cells treated with L. cornuta compared to the negative control (DMSO 0.2%). Conclusion: The findings indicate that L. cornuta ethyl acetate fraction phytochemicals modulates various molecular targets and pathways to exhibit selective antiproliferative and cytotoxic effects against HeLa-229 cells. This study lays a foundation for further research to develop innovative clinical anticervical cancer agents.Copyright © 2024 Lagu, Nyamai and Njeru.