Qin Yan, Gao Ying, Wu Dan, Liu Qing-Qing, Su Chang, Liu Guan, Yang Le, Zhao Ming-Gao, Yao Jing-Yue
Precision Pharmacy and Drug Development Center, Department of Pharmacy, Tangdu Hospital, Air Force Medical University, Xi'an, Shaanxi, China.
Front Pharmacol. 2024 Dec 11;15:1509482. doi: 10.3389/fphar.2024.1509482. eCollection 2024.
Hepatocellular carcinoma (HCC), the third leading cancer mortality worldwide, shows rising incidence. The mitochondria in HCC cells are prone to damage from metabolic stress and oxidative stress, necessitating heightened mitophagy for mitochondrial homeostasis and cell survival. Thus, mitophagy inhibition is a promising HCC therapy. The traditional Chinese medicinal herb ailanthone have proved promote mitochondrial dysfunction and inhibits HCC. However, the underlying mechanism remains unclear.
CCK8 assay was applied to detect the proliferation. JC-1, MitoTracker Red/Green and MitoSOX staining were applied to detect the mitochondrial homeostasis. Inflammatory factors were analysed via ELISA and WB assay. Mitochondria and cytoplasm separation, genome extraction and qPCR were used to detect mitochondrial DNA (mtDNA) leakage. Mitochondria ultrastructure was detected by transmission electron microscopy. WB and IHC experiments were applied to detect protein expression. Protein-protein interactions detected by immunoprecipitation and immunofluorescence imaging. The antitumor effect was validated by the xenograft mouse model.
In this study, we demonstrated the potent anti-HCC properties of ailanthone and revealed its molecular mechanism. studies demonstrated that ailanthone effectively inhibited PINK1-PRKN mediated mitophagy and promoted BAX-BAK1 mitochondrial pores formation through PRKN inhibition. This process led to the mitochondrial mtDNA leakage into the cytoplasm, which subsequently triggered the induction of inflammatory factors. The inhibition of mitophagy and the activation of inflammatory response ultimately led to HCC proliferation inhibition. In vivo studies demonstrated that ailanthone exhibited stronger anti-HCC activity than 5-Fluorouracil (5-FU), with no significant adverse effects on animal body weight or the physiological functions of vital organs.
This study highlighted the efficacy of ailanthone against HCC and elucidated its underlying molecular mechanisms, suggesting the promising therapeutic potential of ailanthone for HCC.
肝细胞癌（HCC）是全球第三大致癌死亡原因，其发病率呈上升趋势。HCC细胞中的线粒体容易受到代谢应激和氧化应激的损伤，因此需要增强线粒体自噬以维持线粒体稳态和细胞存活。因此，抑制线粒体自噬是一种有前景的HCC治疗方法。传统中药臭椿酮已被证明可促进线粒体功能障碍并抑制HCC。然而，其潜在机制仍不清楚。
采用CCK8法检测细胞增殖。采用JC-1、MitoTracker Red/Green和MitoSOX染色检测线粒体稳态。通过ELISA和WB法分析炎症因子。采用线粒体与细胞质分离、基因组提取和qPCR检测线粒体DNA（mtDNA）泄漏。通过透射电子显微镜检测线粒体超微结构。采用WB和IHC实验检测蛋白表达。通过免疫沉淀和免疫荧光成像检测蛋白质-蛋白质相互作用。通过异种移植小鼠模型验证抗肿瘤作用。
在本研究中，我们证明了臭椿酮具有强大的抗HCC特性，并揭示了其分子机制。研究表明，臭椿酮通过抑制PRKN有效抑制PINK1-PRKN介导的线粒体自噬，并促进BAX-BAK1线粒体孔形成。这一过程导致线粒体mtDNA泄漏到细胞质中，随后引发炎症因子的诱导。抑制线粒体自噬和激活炎症反应最终导致HCC增殖抑制。体内研究表明，臭椿酮比5-氟尿嘧啶（5-FU）表现出更强的抗HCC活性，对动物体重或重要器官的生理功能没有显著不良影响。
本研究突出了臭椿酮对HCC的疗效，并阐明了其潜在的分子机制，表明臭椿酮在HCC治疗方面具有广阔的潜力。
