Kamau Sally Wambui, Jepkorir Mercy, Kipkoech Gilbert, Lagu Inyani John Lino, Kanda Wesley, Kibunja Susan, Letoluo Rakita, Barmasai Shadrack, Wanyoko Alice, Ruttoh Vincent, Kuria James, Mwitari Peter Githaiga, Ngugi Mathew Piero, Njeru Sospeter Ngoci
Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Nairobi, Kenya.
Centre for Traditional Medicine and Drug Research (CTMDR), Kenya Medical Research Institute (KEMRI), Nairobi, Kenya.
PLoS One. 2025 Sep 24;20(9):e0331649. doi: 10.1371/journal.pone.0331649. eCollection 2025.
Grewia villosa is a plant native to Kenya, with a traditional history among Ambeere people for treating and managing prostate and breast cancers. Previous scientific studies have demonstrated its anti-inflammatory and antioxidant properties. However, a scientific gap exists on the bioactivity of G. villosa against cervical cancer, particularly on in vitro HeLa cell line model. Additionally, the specific molecular targets and mode of antiproliferative action have not been well elucidated. Therefore, this study sought to investigate the antiproliferative activity, putative targets and mode of action of G. villosa using in vitro cell culture, molecular biology and in silico-based approaches. Antiproliferative analyses were evaluated through MTT assay, cell migration inhibition through in vitro scratch assay, and phytochemical profiling through Gas chromatography-mass spectrometry (GC-MS) analysis. Further, putative targets were identified through network pharmacology approach, computationally validated by molecular docking, and functionally through the real-time quantitative polymerase chain reaction (RT-qPCR) method. The G. villosa ethyl acetate (GVEA) extract fraction was the most active extract fraction, with IC50 of 100.7 µg/mL and a selectivity index of 2.38. Dodecan-2-ylbenzene and 2,6,10-trimethyltetradecane compounds were some notable compounds that can partly be associated with reported antiproliferative activity as they demonstrated strong binding affinity to identified putative targets, including EGFR and AKT1. RT-qPCR analysis functionally confirmed the downregulation of EGFR and AKT1, and the upregulation of tumor protein 53 and Caspase 3 molecular targets, suggesting that GVEA extract indeed perturbs the predicted molecular targets. This study therefore reports the selective antiproliferative properties of the G. villosa ethyl acetate extract fraction in a cervical cancer model (HeLa) cell line while at the same time providing putative targets, which is important in shedding light on potential mechanistic basis of its demonstrated antiproliferative activity. This highlights the plant's potential in discovering products and compounds for further investigation on possible application in cervical cancer management and/or treatment.
绒毛扁担杆是一种原产于肯尼亚的植物，在安贝雷族中有着治疗和管理前列腺癌及乳腺癌的传统历史。先前的科学研究已证明其具有抗炎和抗氧化特性。然而，关于绒毛扁担杆对宫颈癌的生物活性，特别是在体外人宫颈癌细胞系模型方面，存在科学空白。此外，其具体的分子靶点和抗增殖作用模式尚未得到充分阐明。因此，本研究旨在利用体外细胞培养、分子生物学和基于计算机模拟的方法，研究绒毛扁担杆的抗增殖活性、推定靶点及作用模式。通过MTT法评估抗增殖分析，通过体外划痕试验评估细胞迁移抑制，通过气相色谱 - 质谱联用（GC - MS）分析进行植物化学分析。此外，通过网络药理学方法鉴定推定靶点，通过分子对接进行计算验证，并通过实时定量聚合酶链反应（RT - qPCR）方法进行功能验证。绒毛扁担杆乙酸乙酯（GVEA）提取物组分是活性最高的提取物组分，IC50为100.7μg/mL，选择性指数为2.38。十二烷基苯和2,6,10 - 三甲基十四烷化合物是一些值得注意的化合物，它们对已鉴定的推定靶点（包括表皮生长因子受体（EGFR）和蛋白激酶B（AKT1））表现出强结合亲和力，这在一定程度上可与报道的抗增殖活性相关。RT - qPCR分析在功能上证实了EGFR和AKT1的下调以及肿瘤蛋白53和半胱天冬酶3分子靶点的上调，表明GVEA提取物确实干扰了预测的分子靶点。因此，本研究报告了绒毛扁担杆乙酸乙酯提取物组分在宫颈癌模型（HeLa）细胞系中的选择性抗增殖特性，同时提供了推定靶点，这对于阐明其已证明的抗增殖活性的潜在机制基础具有重要意义。这突出了该植物在发现用于进一步研究宫颈癌管理和/或治疗中可能应用的产品和化合物方面的潜力。
