Untargeted metabolomics using UHPLC-Q-Orbitrap HRMS for identifying cytotoxic compounds on MCF-7 breast cancer cells from Annona muricata Linn leaf extracts as potential anticancer agents.

IF 3 3区生物学 Q2 BIOCHEMICAL RESEARCH METHODS Phytochemical Analysis Pub Date : 2024-08-01 Epub Date: 2024-05-06 DOI:10.1002/pca.3373

Dewi Anggraini Septaningsih, Irma Herawati Suparto, Suminar Setiati Achmadi, Rudi Heryanto, Mohamad Rafi

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Abstract

Introduction: The leaves of Annona muricata L., known as "soursop" or "sirsak" in Indonesia, are used traditionally for cancer treatment. However, the bioactive components remain largely unidentified.

Objective: This study used untargeted liquid chromatography-tandem mass spectrometry (LC-MS/MS)-based metabolomics to identify potential cytotoxic compounds in A. muricata leaf extracts on MCF-7 breast cancer cells in vitro.

Methods: A. muricata leaves were macerated with water, 99% ethanol, and aqueous mixtures containing 30%, 50%, and 80% ethanol. Cytotoxic activity of the extracts against MCF-7 breast cancer cells was determined using the MTT assay. Ultra-high-performance liquid chromatography-Q-Orbitrap high-resolution mass spectroscopy (UHPLC-Q-Orbitrap-HRMS) was used to characterize the metabolite composition of each extract. The correlations between metabolite profile and cytotoxic activities were evaluated using orthogonal partial least square discriminant analysis (OPLS-DA). The binding of these bioactive compounds to the tumorigenic alpha-estrogen receptor (3ERT) was then evaluated by in silico docking simulations.

Results: Ninety-nine percent ethanol extracts demonstrated the greatest potency for reducing MCF-7 cell viability (IC₅₀ = 22 μg/ml). We detected 35 metabolites in ethanol extracts, including alkaloids, flavonoids, and acetogenins. OPLS-DA predicted that annoreticuin, squadiolin C, and xylopine, and six unknown acetogenin metabolites, might reduce MCF-7 cell viability. In silico analysis predicted that annoreticuin, squadiolin C, and xylopine bind to 3ERT with an affinity comparable to doxorubicin.

Conclusion: Untargeted metabolomics and in silico modeling identified cytotoxic compounds on MCF-7 cells and binding affinity to 3ERT in A. muricata leaf extracts. The findings need to be further verified to prove the screening results.

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利用 UHPLC-Q-Orbitrap HRMS 进行非靶向代谢组学研究，从 Annona muricata Linn 叶提取物中鉴定对 MCF-7 乳腺癌细胞具有细胞毒性的化合物，并将其作为潜在的抗癌剂。

简介Annona muricata L.在印度尼西亚被称为 "soursop "或 "sirsak"，其叶子在传统上被用于治疗癌症。然而，其中的生物活性成分在很大程度上仍未被发现：本研究采用基于非靶向液相色谱-串联质谱（LC-MS/MS）的代谢组学方法，鉴定A. muricata叶提取物中对体外MCF-7乳腺癌细胞具有潜在细胞毒性的化合物：方法：用清水、99% 的乙醇和含 30%、50% 和 80% 乙醇的水混合物浸泡 A. muricata 叶子。采用 MTT 法测定提取物对 MCF-7 乳腺癌细胞的细胞毒活性。超高效液相色谱-Q-Orbitrap 高分辨率质谱（UHPLC-Q-Orbitrap-HRMS）用于表征每种提取物的代谢物组成。采用正交偏最小二乘法判别分析（OPLS-DA）评估了代谢物特征与细胞毒性活性之间的相关性。然后通过硅对接模拟评估了这些生物活性化合物与肿瘤α-雌激素受体（3ERT）的结合情况：结果：99% 的乙醇提取物在降低 MCF-7 细胞活力方面表现出最大的效力（IC50 = 22 μg/ml）。我们在乙醇提取物中检测到 35 种代谢物，包括生物碱、黄酮类化合物和乙酰苷元。根据 OPLS-DA 预测，胭脂虫素、番茄红素 C 和木犀草素以及六种未知的苷元代谢物可能会降低 MCF-7 细胞的活力。硅学分析预测，annoreticuin、squadiolin C 和 xylopine 与 3ERT 的结合亲和力与多柔比星相当：结论：非靶向代谢组学和硅学建模发现了A. muricata叶提取物中对MCF-7细胞具有细胞毒性的化合物以及与3ERT的结合亲和力。这些发现需要进一步验证，以证明筛选结果。

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来源期刊

Phytochemical Analysis 生物-分析化学

CiteScore

6.00

自引率

6.10%

发文量

审稿时长

1.7 months

期刊介绍： Phytochemical Analysis is devoted to the publication of original articles concerning the development, improvement, validation and/or extension of application of analytical methodology in the plant sciences. The spectrum of coverage is broad, encompassing methods and techniques relevant to the detection (including bio-screening), extraction, separation, purification, identification and quantification of compounds in plant biochemistry, plant cellular and molecular biology, plant biotechnology, the food sciences, agriculture and horticulture. The Journal publishes papers describing significant novelty in the analysis of whole plants (including algae), plant cells, tissues and organs, plant-derived extracts and plant products (including those which have been partially or completely refined for use in the food, agrochemical, pharmaceutical and related industries). All forms of physical, chemical, biochemical, spectroscopic, radiometric, electrometric, chromatographic, metabolomic and chemometric investigations of plant products (monomeric species as well as polymeric molecules such as nucleic acids, proteins, lipids and carbohydrates) are included within the remit of the Journal. Papers dealing with novel methods relating to areas such as data handling/ data mining in plant sciences will also be welcomed.