CaMKII Exacerbates Doxorubicin-Induced Cardiotoxicity by Promoting Ubiquitination Through USP10 Inhibition

IF 2.9 2区医学 Q2 ONCOLOGY

Cancer Medicine Pub Date : 2024-11-08 DOI:10.1002/cam4.70286

Yitong Yang, Zhenyi Wang, Nisha Wang, Jian Yang, Lifang Yang

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Abstract

Background

Doxorubicin (DOX) is an effective anticancer drug, but it has a problem of cardiotoxicity that cannot be ignored. Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) is tightly associated with the pathological progression of DOX-induced cardiotoxicity. Ubiquitin-specific protease 10 (USP10) plays an important role in many biological processes and cancers. However, its association with DOX-induced cardiotoxicity and CaMKII remains unclear.

Methods

H9C2 cells, HL-1 cells and C57BL/6 mice were used to establish the DOX-induced cardiotoxicity model, and the CaMKII-specific inhibitor KN-93 and USP10 specific inhibitor Spautin-1 were used to observe the CaMKII and USP10 effect. In cell experiments, CCK-8 method was used to assess cell viability, LDH kit was used to assess lactate dehydrogenase expression, DCFH-DA staining was used to observe changes in active oxygen content, TUNEL staining was used to observe cell apoptosis, and Western blotting method was used to detect relevant protein markers. The expression of p-CaMKII and USP10 was assessed by immunofluorescence staining. In animal experiments, mouse echocardiograph was used were used to evaluate cardiac function, and HE staining and Masson staining were used to evaluate myocardial injury. Cardiomyocyte apoptosis was detected by TUNEL staining. Western blotting method was used to detect relevant protein markers.

Results

Our results demonstrated that activation of CaMKII and inhibition of USP10 pathway related to DOX-induced cardiotoxicity. Inhibition of CaMKII with KN-93 ameliorated DOX-induced cardiac dysfunction and cytotoxicity. In addition, CaMKII inhibition prevented DOX-induced apoptosis and ubiquitination. Furthermore, CaMKII inhibition increased USP10 expression in DOX-treated mouse hearts, H9C2 cells and HL-1 cells. At last, the USP10 inhibitor, Spautin-1, blocked the regulatory effect of CaMKII inhibition on apoptosis and ubiquitination in DOX-induced cardiotoxicity.

Conclusion

Our findings revealed that DOX-induced myocardial apoptosis and activated CaMKII through cellular and animal levels, while providing a novel probe into the mechanism of CaMKII action: promoting ubiquitination by inhibiting USP10 aggravated apoptosis.

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CaMKII通过抑制USP10促进泛素化而加剧多柔比星诱导的心脏毒性

背景：多柔比星（DOX）是一种有效的抗癌药物，但其心脏毒性问题不容忽视。Ca2+/钙调蛋白依赖性蛋白激酶II（CaMKII）与DOX诱导的心脏毒性的病理进展密切相关。泛素特异性蛋白酶 10（USP10）在许多生物过程和癌症中发挥着重要作用。方法：用 H9C2 细胞、HL-1 细胞和 C57BL/6 小鼠建立 DOX 诱导的心脏毒性模型，用 CaMKII 特异性抑制剂 KN-93 和 USP10 特异性抑制剂 Spautin-1 观察 CaMKII 和 USP10 的作用。在细胞实验中，使用 CCK-8 法评估细胞活力，使用 LDH 试剂盒评估乳酸脱氢酶的表达，使用 DCFH-DA 染色法观察活性氧含量的变化，使用 TUNEL 染色法观察细胞凋亡，使用 Western 印迹法检测相关蛋白标记物。免疫荧光染色法评估了 p-CaMKII 和 USP10 的表达。在动物实验中，用小鼠超声心动图评估心脏功能，用 HE 染色和 Masson 染色评估心肌损伤。通过 TUNEL 染色检测心肌细胞凋亡。采用 Western 印迹法检测相关蛋白标记物：结果：我们的研究结果表明，CaMKII的激活和USP10通路的抑制与DOX诱导的心脏毒性有关。用 KN-93 抑制 CaMKII 可改善 DOX 诱导的心脏功能障碍和细胞毒性。此外，抑制 CaMKII 还能防止 DOX 诱导的细胞凋亡和泛素化。此外，抑制 CaMKII 还能增加 USP10 在 DOX 处理的小鼠心脏、H9C2 细胞和 HL-1 细胞中的表达。最后，USP10抑制剂Spautin-1阻断了CaMKII抑制在DOX诱导的心脏毒性中对细胞凋亡和泛素化的调节作用：我们的研究结果表明，DOX诱导心肌细胞凋亡，并通过细胞和动物水平激活CaMKII，同时为CaMKII的作用机制提供了一个新的探究方法：通过抑制USP10促进泛素化，从而加剧细胞凋亡。

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来源期刊

Cancer Medicine ONCOLOGY-

CiteScore

5.50

自引率

2.50%

发文量

907

审稿时长

19 weeks

期刊介绍： Cancer Medicine is a peer-reviewed, open access, interdisciplinary journal providing rapid publication of research from global biomedical researchers across the cancer sciences. The journal will consider submissions from all oncologic specialties, including, but not limited to, the following areas: Clinical Cancer Research Translational research ∙ clinical trials ∙ chemotherapy ∙ radiation therapy ∙ surgical therapy ∙ clinical observations ∙ clinical guidelines ∙ genetic consultation ∙ ethical considerations Cancer Biology: Molecular biology ∙ cellular biology ∙ molecular genetics ∙ genomics ∙ immunology ∙ epigenetics ∙ metabolic studies ∙ proteomics ∙ cytopathology ∙ carcinogenesis ∙ drug discovery and delivery. Cancer Prevention: Behavioral science ∙ psychosocial studies ∙ screening ∙ nutrition ∙ epidemiology and prevention ∙ community outreach. Bioinformatics: Gene expressions profiles ∙ gene regulation networks ∙ genome bioinformatics ∙ pathwayanalysis ∙ prognostic biomarkers. Cancer Medicine publishes original research articles, systematic reviews, meta-analyses, and research methods papers, along with invited editorials and commentaries. Original research papers must report well-conducted research with conclusions supported by the data presented in the paper.