Dynorphin B induces mitochondrial fragmentation in NSCLC through the PKD/DRP-1 signaling pathway

IF 2.5 3区医学 Q3 ENDOCRINOLOGY & METABOLISM

Neuropeptides Pub Date : 2025-06-25 DOI:10.1016/j.npep.2025.102535

Yunxiao Li , Bin Zhou , Yuenan Yang , Kexin Liu , Shichao Zhou

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引用次数: 0

Abstract

Mitochondrial fragmentation and impairment are essential targets for therapeutic approach for non-small cell lung cancer (NSCLC), given their significant contributions to the persistence and progression of malignant cells. Dynorphin B (Dyn B), an endogenous opioid peptide, has been demonstrated for its involvement in an extensive array of cellular activities; however, its specific functions and mechanisms within the context of cancer remain largely undefined. To address this, we employed NCI-H2087 NSCLC cells treated with Dyn B (0.01–100 μM) and utilized lactate dehydrogenase (LDH) release and γ-glutamyl transpeptidase (GPT) activity assays to evaluate cytotoxicity. Mitochondrial function was assessed via Complex I activity assays, adenosine triphosphate (ATP) production measurements, and MitoSOX Green staining for reactive oxygen species (ROS). MitoTracker Red staining with ImageJ quantification characterized mitochondrial morphology, while Western blot analysis probed phosphorylation of dynamin-related protein 1 (DRP1) and protein kinase D (PKD). Lentiviral shRNA-mediated PKD silencing was used to validate functional rescue of mitochondrial dynamics. This investigation reveals that Dyn B induces cytotoxic effects in NCI-H2087 NSCLC cells by facilitating mitochondrial dysfunction and fragmentation. Treatment with Dyn B resulted in a significant augmentation of LDH and elevated GPT activity, indicating cellular injury. Additionally, Dyn B compromised mitochondrial functionality by reducing Complex I activity, diminishing ATP synthesis, and promoting mitochondrial ROS generation. Mechanistically, Dyn B triggered mitochondrial fragmentation through activation of DRP1 and PKD, without affecting protein kinase C (PKC). Silencing of PKD reversed Dyn B–induced mitochondrial fragmentation and restored mitochondrial functionality. These findings underscore the promising role of Dyn B as a prospective therapeutic agent in NSCLC, targeting mitochondrial dynamics via the PKD-DRP1 signaling pathway.

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Dynorphin B通过PKD/DRP-1信号通路诱导NSCLC线粒体断裂

线粒体断裂和损伤是治疗非小细胞肺癌（NSCLC）的重要靶点，因为它们对恶性细胞的持续和进展有重要贡献。肌啡肽B （Dyn B）是一种内源性阿片肽，已被证明参与广泛的细胞活动；然而，其在癌症中的具体功能和机制在很大程度上仍未明确。为了解决这个问题，我们使用Dyn B （0.01-100 μM）处理NCI-H2087 NSCLC细胞，并利用乳酸脱氢酶（LDH）释放和γ-谷氨酰转肽酶（GPT）活性测定来评估细胞毒性。通过复合体I活性测定、三磷酸腺苷（ATP）生成测定和MitoSOX绿色活性氧（ROS）染色来评估线粒体功能。MitoTracker Red用ImageJ定量染色表征线粒体形态，Western blot分析检测动力蛋白相关蛋白1 （DRP1）和蛋白激酶D （PKD）的磷酸化。慢病毒shrna介导的PKD沉默被用来验证线粒体动力学的功能拯救。本研究表明，Dyn B通过促进线粒体功能障碍和断裂，诱导NCI-H2087 NSCLC细胞的细胞毒性作用。Dyn B治疗导致LDH显著增加，GPT活性升高，表明细胞损伤。此外，Dyn B通过降低复合物I活性、减少ATP合成和促进线粒体ROS生成来损害线粒体功能。在机制上，Dyn B通过激活DRP1和PKD触发线粒体断裂，而不影响蛋白激酶C （PKC）。PKD的沉默逆转了Dyn b诱导的线粒体断裂并恢复了线粒体功能。这些发现强调了Dyn B作为一种有前景的治疗药物在NSCLC中的作用，通过PKD-DRP1信号通路靶向线粒体动力学。

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来源期刊

Neuropeptides 医学-内分泌学与代谢

CiteScore

5.40

自引率

6.90%

发文量

审稿时长

>12 weeks

期刊介绍： The aim of Neuropeptides is the rapid publication of original research and review articles, dealing with the structure, distribution, actions and functions of peptides in the central and peripheral nervous systems. The explosion of research activity in this field has led to the identification of numerous naturally occurring endogenous peptides which act as neurotransmitters, neuromodulators, or trophic factors, to mediate nervous system functions. Increasing numbers of non-peptide ligands of neuropeptide receptors have been developed, which act as agonists or antagonists in peptidergic systems. The journal provides a unique opportunity of integrating the many disciplines involved in all neuropeptide research. The journal publishes articles on all aspects of the neuropeptide field, with particular emphasis on gene regulation of peptide expression, peptide receptor subtypes, transgenic and knockout mice with mutations in genes for neuropeptides and peptide receptors, neuroanatomy, physiology, behaviour, neurotrophic factors, preclinical drug evaluation, clinical studies, and clinical trials.