乙酸乙酯提取物对宫颈癌细胞抗增殖作用的网络药理学、分子对接及实验方法
Network pharmacology, molecular docking and experimental approaches of the anti-proliferative effects of ethyl-acetate extract in cervical cancer cells.
作者信息
Kamau Sally Wambui, Ngugi Mathew Piero, Mwitari Peter Githaiga, Njeru Sospeter Ngoci
机构信息
Center for Traditional Medicine and Drug Research, Kenya Medical Research Institute, Kenya.
Department of Biochemistry, Microbiology and Biotechnology, Kenyatta University, Kenya.
出版信息
Heliyon. 2024 Sep 2;10(17):e37324. doi: 10.1016/j.heliyon.2024.e37324. eCollection 2024 Sep 15.
BACKGROUND
Cervical cancer, one of the lethal cancers among women, is a challenging disease to treat. The current therapies often come with severe side effects and the risk of resistance development. Traditional herbal medicine, with its potential to offer effective and less toxic options, is a promising avenue. This study was undertaken to investigate the potential of () root bark extracts in selectively inhibiting the proliferation of cervical cancer cells, using the HeLa cell line as an model.
METHODS
plant extracts were first screened at a fixed concentration of 200 μg/ml to determine the active extract. The selective anti-proliferative activity of the active extract was evaluated in a concentration dilution assay using the (3-(4,5-dimethylthiazol- 2-yl)-2,5-diphenyltetrazolium bromide) MTT assay on cancerous (HeLa) cells and non-cancerous (Vero) cells to determine the half-maximal inhibitory (IC) and half-cytotoxic concentrations (CC), respectively. Functional assays on cell morphology (by microscopy), cell migration (wound healing assay) and cell cycle (by flow cytometry) were also conducted. The active extract was analyzed using Gas Chromatography/Mass Spectrometry (GC/MS) to determine any compounds it contained. Following identification of possible gene targets by network pharmacology, the genes were validated by molecular docking and Real-Time Quantitative Polymerase Chain Reaction (RT-qPCR).
RESULTS
The ethyl acetate extract of (EARP), the most active extract, selectively inhibited the growth of cervical cancer cells, their migration and induced cell cycle arrest at the S phase. analysis revealed that squalene, 3,3a,6,6-tetramethyl-4,5,5a,7,8,9-hexahydro-1H-cyclopenta[i]indene and Olean-12-en-3.beta.-ol, acetate showed acceptable drug-like characteristics and may be partly attributed to the bioactivity demonstrated and the deregulation of the mRNA expression of AKT1, NF-κB, p53, Bax, Bcl-2, and Er-b-B2.
CONCLUSION
This study, for the first time, demonstrates the anti-proliferation effects of EARP and forms a firm foundation for further drug development studies.
背景
宫颈癌是女性致死性癌症之一，是一种具有挑战性的疾病。目前的治疗方法往往伴有严重的副作用和产生耐药性的风险。传统草药有提供有效且毒性较小选择的潜力，是一条有前景的途径。本研究旨在以HeLa细胞系为模型，研究（）根皮提取物选择性抑制宫颈癌细胞增殖的潜力。
方法
首先以200μg/ml的固定浓度筛选植物提取物以确定活性提取物。使用（3-（4,5-二甲基噻唑-2-基）-2,5-二苯基四氮唑溴盐）MTT法，在浓度稀释试验中评估活性提取物对癌细胞（HeLa）和非癌细胞（Vero）的选择性抗增殖活性，分别确定半数最大抑制浓度（IC）和半数细胞毒性浓度（CC）。还进行了细胞形态（通过显微镜）、细胞迁移（伤口愈合试验）和细胞周期（通过流式细胞术）的功能试验。使用气相色谱/质谱联用仪（GC/MS）分析活性提取物以确定其所含的任何化合物。通过网络药理学鉴定可能的基因靶点后，通过分子对接和实时定量聚合酶链反应（RT-qPCR）对基因进行验证。
结果
活性最强的提取物——（）乙酸乙酯提取物（EARP）选择性抑制宫颈癌细胞的生长、迁移，并诱导细胞周期停滞于S期。分析表明，角鲨烯、3,3a,6,6-四甲基-4,5,5a,7,8,9-六氢-1H-环戊并[i]茚和齐墩果-12-烯-3β-醇乙酸酯具有可接受的类药物特性，这可能部分归因于所表现出的生物活性以及AKT1、NF-κB、p53、Bax、Bcl-2和Er-b-B2的mRNA表达失调。
结论
本研究首次证明了EARP的抗增殖作用，为进一步的药物开发研究奠定了坚实基础。
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