Bioinformatics-Guided Discovery of miRNAs Involved in Apoptosis Modulated by Parthenolide Combined with Vincristine in The NALM6 Cell Line.

IF 16.4 1区化学 Q1 CHEMISTRY, MULTIDISCIPLINARY

Accounts of Chemical Research Pub Date : 2024-02-01 DOI:10.22074/cellj.2024.2013673.1428

Atefeh Bahmei, Sepideh Namdari, Mohammad Yaghoubzad-Maleki, Ali Emami, Reza Ranjbaran, Gholamhossein Tamaddon

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引用次数: 0

Abstract

Objective: Acute lymphoblastic leukemia (ALL) is a highly heterogeneous leukemia. Despite the current improvement in conventional chemotherapy and high survival rates, the outcomes remain challenging. Sesquiterpen extracted from the Tanacetum parthenium, parthenolide, is a potential anticancer agent that can modulate the expression of miRNAs and induce apoptosis. The objective of this study was to investigate the effect of parthenolide in combination with vincristine and alone on the apoptosis rate and expression of miR-125b-5p, miR-181b-5p, and miR-17-5p in the NALM6 cell line.

Materials and methods: In this experimental study, cell viability and metabolic activity were determined through MTT assay and PI staining. Flow cytometry was applied to evaluate the rate of apoptosis. The expression of miRNAs was assessed using real-time polymerase chain reaction. Bioinformatic analyses, including Cytoscape, RNAhybrid, and signaling pathway analysis were employed to investigate the association of miR-17-5p, miR-181b-5p and miR-125b- 5p with apoptosis. Further, molecular docking served to validate the modulation of these miRNAs by parthenolide and vincristine treatment.

Results: The MTT assay indicated that 7.7 μM of parthenolide decreased the metabolic activity to 50% after 48 hours. PI staining analysis indicated that at concentrations below the half maximal inhibitory concentration, parthenolide caused 50% cell death. Flow cytometric analysis indicated that parthenolide (1.925 μM) in combination with vincristine (1.2 nM) induced apoptosis in 83.2% of the cells. Real-time quantitative reverse transcription polymerase chain reaction (qRTPCR) analysis showed significant changes in the expression levels of miR-17-5p, miR-125b-5p, and miR-181b-5p. Moreover, the combination therapy downregulated the expression of miRNAs significantly. This was consistent with our bioinformatic analysis demonstrating that the studied miRNAs are regulators of apoptosis. Finally, molecular docking validated the modulation of the miRNAs by parthenolide and vincristine.

Conclusion: Parthenolide in combination with vincristine triggers apoptosis at a high rate in the NALM6 cell line. Moreover, this combination therapy can decrease the expression of miR-17-5p, miR-181b-5p, and miR-125b-5p.

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生物信息学指导下发现NALM6细胞系中参与帕替诺利与长春新碱联合调控细胞凋亡的miRNAs

目的：急性淋巴细胞白血病（ALL）是一种高度异质性的白血病：急性淋巴细胞白血病（ALL）是一种高度异质性白血病。尽管目前常规化疗的疗效有所改善，患者的存活率也很高，但其治疗结果仍具有挑战性。从丹参（Tanacetum parthenium）中提取的半萜--丹参内酯（parthenolide）是一种潜在的抗癌剂，可调节 miRNAs 的表达并诱导细胞凋亡。本研究的目的是探讨马钱子内酯与长春新碱联合或单独使用对 NALM6 细胞系的凋亡率及 miR-125b-5p、miR-181b-5p 和 miR-17-5p 表达的影响：在本实验研究中，通过 MTT 试验和 PI 染色测定细胞活力和代谢活性。流式细胞术用于评估细胞凋亡率。使用实时聚合酶链反应评估 miRNA 的表达。生物信息学分析包括 Cytoscape、RNAhybrid 和信号通路分析，用于研究 miR-17-5p、miR-181b-5p 和 miR-125b- 5p 与细胞凋亡的关系。此外，分子对接法还验证了马钱子内酯和长春新碱对这些 miRNAs 的调控作用：MTT 试验表明，7.7 μM 的马钱子内酯可在 48 小时后将代谢活性降至 50%。PI 染色分析表明，当浓度低于半数最大抑制浓度时，非那西丁内酯会导致 50%的细胞死亡。流式细胞分析表明，非那西丁内酯（1.925 μM）与长春新碱（1.2 nM）联用可诱导 83.2% 的细胞凋亡。实时定量反转录聚合酶链反应（qRTPCR）分析显示，miR-17-5p、miR-125b-5p 和 miR-181b-5p 的表达水平发生了显著变化。此外，联合疗法显著下调了 miRNA 的表达。这与我们的生物信息学分析表明所研究的 miRNAs 是细胞凋亡的调控因子是一致的。最后，分子对接验证了parthenolide和长春新碱对miRNAs的调节作用：结论：帕菲诺莱与长春新碱联用可诱导 NALM6 细胞系高度凋亡。此外，这种联合疗法还能降低 miR-17-5p、miR-181b-5p 和 miR-125b-5p 的表达。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Accounts of Chemical Research 化学-化学综合

CiteScore

31.40

自引率

1.10%

发文量

312

审稿时长

2 months

期刊介绍： Accounts of Chemical Research presents short, concise and critical articles offering easy-to-read overviews of basic research and applications in all areas of chemistry and biochemistry. These short reviews focus on research from the author’s own laboratory and are designed to teach the reader about a research project. In addition, Accounts of Chemical Research publishes commentaries that give an informed opinion on a current research problem. Special Issues online are devoted to a single topic of unusual activity and significance. Accounts of Chemical Research replaces the traditional article abstract with an article "Conspectus." These entries synopsize the research affording the reader a closer look at the content and significance of an article. Through this provision of a more detailed description of the article contents, the Conspectus enhances the article's discoverability by search engines and the exposure for the research.