Morusin, a novel inhibitor of ACLY, induces mitochondrial apoptosis in hepatocellular carcinoma cells through ROS-mediated mitophagy

IF 6.9 2区医学 Q1 MEDICINE, RESEARCH & EXPERIMENTAL

Biomedicine & Pharmacotherapy Pub Date : 2024-09-27 DOI:10.1016/j.biopha.2024.117510

Desheng Li , Xiaoqing Yuan , Jianjun Ma , Tao Lu , Jinjin Zhang , Huan Liu , Guanqing Zhang , Yue Wang , Xiaohan Liu , Qiqiang Xie , Ling Zhou , Maolei Xu

{"title":"Morusin, a novel inhibitor of ACLY, induces mitochondrial apoptosis in hepatocellular carcinoma cells through ROS-mediated mitophagy","authors":"Desheng Li , Xiaoqing Yuan , Jianjun Ma , Tao Lu , Jinjin Zhang , Huan Liu , Guanqing Zhang , Yue Wang , Xiaohan Liu , Qiqiang Xie , Ling Zhou , Maolei Xu","doi":"10.1016/j.biopha.2024.117510","DOIUrl":null,"url":null,"abstract":"<div><h3>Objective</h3><div>Morusin (Mor), a prenylated flavonoid isolated from the root bark of <em>Morus alba L</em>., exhibits potent anti-tumour effects; however, the molecular target of Mor is still not entirely clear. This study aimed to elucidate the mechanism of Mor against hepatocellular carcinoma (HCC) and identify potential molecular targets.</div></div><div><h3>Methods</h3><div>Mitochondrial function was assessed by measuring the mitochondrial membrane potential, mitochondrial ultrastructure, oxygen consumption, and ATP levels. Mor-induced mitophagy was confirmed using western blotting, immunofluorescence, and fluorescent probes. Transcriptomics, flow cytometry, western blotting, qRT-PCR and biochemical assays were used to reveal the molecular mechanisms and targets of Mor against HCC. We further validated the interaction between Mor and the target proteins using molecular docking and biolayer interferometry (BLI). The inhibitory effect of Mor <em>in vivo</em> was evaluated using a Hep3B murine xenograft model.</div></div><div><h3>Results</h3><div>Mor significantly reduced the ATP citrate lyase (ACLY) expression and inhibited ACLY activity in HCC cells. BLI analysis demonstrated a direct interaction between Mor and the ACLY active domain. Mor-induced ACLY inhibition led to ROS accumulation in HCC cells, which caused mitochondrial damage, triggered PINK1/Parkin-mediated mitophagy, and ultimately induced mitochondrial apoptosis. We further verified that ROS is crucial in the apoptotic action of Mor through experiments regarding an ROS scavenger. Mor also significantly inhibited tumour xenograft growth <em>in vivo</em>. In addition, analysis of human liver cancer clinical samples revealed elevated ACLY levels positively correlated with histologic grade.</div></div><div><h3>Conclusion</h3><div>Collectively, our findings highlight Mor as a potent bioactive inhibitor of ACLY and a promising candidate for HCC therapy.</div></div>","PeriodicalId":8966,"journal":{"name":"Biomedicine & Pharmacotherapy","volume":"180 ","pages":"Article 117510"},"PeriodicalIF":6.9000,"publicationDate":"2024-09-27","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biomedicine & Pharmacotherapy","FirstCategoryId":"3","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0753332224013969","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}

引用次数: 0

Abstract

Objective

Morusin (Mor), a prenylated flavonoid isolated from the root bark of Morus alba L., exhibits potent anti-tumour effects; however, the molecular target of Mor is still not entirely clear. This study aimed to elucidate the mechanism of Mor against hepatocellular carcinoma (HCC) and identify potential molecular targets.

Methods

Mitochondrial function was assessed by measuring the mitochondrial membrane potential, mitochondrial ultrastructure, oxygen consumption, and ATP levels. Mor-induced mitophagy was confirmed using western blotting, immunofluorescence, and fluorescent probes. Transcriptomics, flow cytometry, western blotting, qRT-PCR and biochemical assays were used to reveal the molecular mechanisms and targets of Mor against HCC. We further validated the interaction between Mor and the target proteins using molecular docking and biolayer interferometry (BLI). The inhibitory effect of Mor in vivo was evaluated using a Hep3B murine xenograft model.

Results

Mor significantly reduced the ATP citrate lyase (ACLY) expression and inhibited ACLY activity in HCC cells. BLI analysis demonstrated a direct interaction between Mor and the ACLY active domain. Mor-induced ACLY inhibition led to ROS accumulation in HCC cells, which caused mitochondrial damage, triggered PINK1/Parkin-mediated mitophagy, and ultimately induced mitochondrial apoptosis. We further verified that ROS is crucial in the apoptotic action of Mor through experiments regarding an ROS scavenger. Mor also significantly inhibited tumour xenograft growth in vivo. In addition, analysis of human liver cancer clinical samples revealed elevated ACLY levels positively correlated with histologic grade.

Conclusion

Collectively, our findings highlight Mor as a potent bioactive inhibitor of ACLY and a promising candidate for HCC therapy.

查看原文本刊更多论文

新型 ACLY 抑制剂 Morusin 通过 ROS 介导的有丝分裂诱导肝癌细胞线粒体凋亡

目的Morusin（Mor）是从桑树根皮中分离出来的一种前烯化黄酮类化合物，具有很强的抗肿瘤作用；然而，Mor的分子靶点仍不完全清楚。本研究旨在阐明 Mor 抗肝细胞癌（HCC）的机制，并确定潜在的分子靶点。方法通过测量线粒体膜电位、线粒体超微结构、耗氧量和 ATP 水平来评估线粒体功能。使用 Western 印迹、免疫荧光和荧光探针证实了 Mor 诱导的有丝分裂。我们利用转录组学、流式细胞术、Western 印迹、qRT-PCR 和生化分析揭示了 Mor 抗 HCC 的分子机制和靶点。我们还利用分子对接和生物层干涉仪（BLI）进一步验证了Mor与靶蛋白之间的相互作用。我们使用 Hep3B 小鼠异种移植模型评估了 Mor 在体内的抑制作用。BLI 分析表明 Mor 与 ACLY 活性结构域之间存在直接相互作用。Mor诱导的ACLY抑制导致ROS在HCC细胞中积累，造成线粒体损伤，引发PINK1/Parkin介导的有丝分裂，最终诱导线粒体凋亡。我们通过ROS清除剂的实验进一步验证了ROS在Mor的凋亡作用中的关键作用。Mor 还能明显抑制体内肿瘤异种移植的生长。此外，对人类肝癌临床样本的分析表明，ACLY 水平的升高与组织学分级呈正相关。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

求助全文

约1分钟内获得全文求助全文

来源期刊

Biomedicine & Pharmacotherapy 医学-药学

CiteScore

11.90

自引率

2.70%

发文量

1621

审稿时长

48 days

期刊介绍： Biomedicine & Pharmacotherapy stands as a multidisciplinary journal, presenting a spectrum of original research reports, reviews, and communications in the realms of clinical and basic medicine, as well as pharmacology. The journal spans various fields, including Cancer, Nutriceutics, Neurodegenerative, Cardiac, and Infectious Diseases.