Circ_0001312 Silencing Suppresses Doxorubicin-Induced Cardiotoxicity via MiR-409-3p/HMGB1 Axis

Abstract

Doxorubicin (DOX) is a potent cytotoxic chemotherapeutic agent limited in clinical application owing to its cumulative and irreversible cardiotoxicity. Circ_0001312 is highly expressed in patients with heart failure. However, it is still unclear whether circ_0001312 plays any roles in DOX-induced cardiotoxicity.Human AC16 cardiomyocytes in functional group were stimulated with DOX. The levels of genes and proteins were detected by qRT-PCR and western blotting. The proliferation, apoptosis, as well as inflammatory and oxidative injury in cardiomyocytes were investigated. Dual-luciferase reporter, RNA immunoprecipitation, and pull-down assays were utilized to confirm the binding between miR-409-3p and circ_0001312 or HMGB1 (high-mobility group box 1). Exosomes were isolated by using the commercial kit and identified by transmission electron microscopy (TEM) and nanoparticle-tracking analysis (NTA).DOX impaired cardiomyocyte proliferation and induced apoptotic, inflammatory, and oxidative injury in cells. Furthermore, it promoted circ_0001312 expression, and the knockdown of circ_0001312 could reverse DOX-evoked cardiomyocyte injury. In terms of mechanics, circ_0001312 bound competitively to miR-409-3p to up-regulate HMGB1, which was a target of miR-409-3p. DOX decreased the miR-409-3p but increased the HMGB1 expression in cardiomyocytes. Functionally, miR-409-3p inhibition attenuated the protective action of circ_0001312 silencing on cardiomyocytes under DOX treatment. Moreover, miR-409-3p could abate DOXevoked apoptosis, and inflammation and oxidative stress in cardiomyocytes, and these effects were counteracted by HMGB1 overexpression. In addition, circ_0001312 was secreted by exosomes and could be transmitted via exosomes.Circ_0001312 reversed the cytotoxic effects mediated by DOX on cardiomyocytes via the miR-409-3p/ HMGB1 axis. Besides, it was released to the extracellular space by exosomes.

Clinical Significance of Non-Coding RNA Regulation of Programmed Cell Death in Hepatocellular Carcinoma

“…miR-409-3p can target high-mobility group box 1 (HMGB1), thereby reducing DOX-induced myocardial cell apoptosis. This further confirms the regulatory role of circRNA_0001312 in the process of DOX-induced myocardial cell injury [ 139 ].…”

Identification of circular RNAs regulating cardiomyocyte proliferation in neonatal pig hearts

“…Although we acknowledge limitations such as sample variation and the need for further validation, our findings will greatly enhance our understanding of circRNA function and its potential implications in heart development and maturation in large mammals. It was reported that modulating the expression of circRNAs confers cardioprotection in preclinical animal models of heart failure (19,64,65). In this study, we identified and validated six circRNAs that regulate cardiomyocyte cell cycle and proliferation.…”

Efeito Protetor do RNA Não Codificante Longo OXCT1-AS1 na Apoptose de Células Miocárdicas Humanas Induzida pela Doxorrubicina pelo Padrão Competitivo de RNA Endógeno

“…Human AC16 cardiomyocytes were obtained from ATCC (Rockville, MD, USA) and maintained in Dulbecco's modified Eagle medium (Gibco, Grand Island, NY, USA) supplemented with 10% fetal bovine serum (Gibco) and 1% penicillin-streptomycin (Gibco) in a humidified atmosphere at 37°C with 95% air and 5% CO 2 . To induce a myocardial cell injury model, AC16 cells were treated with 5 μM DOX (Sangon Biotech Co., Ltd., Shanghai, China) for 24 h. 17 In some experiments, AC16 cells were treated with 5 μM DOX for different periods (0, 0.5, 2, 6, 12, 24, and 48 h) for the detection of RNA expression.…”

“…Para induzir um modelo de lesão celular miocárdica, as células AC16 foram tratadas com 5 μM de DOX (Sangon Biotech Co., Ltd., Xangai, China) por 24 horas. 17 Em alguns experimentos, as células AC16 foram tratadas com 5 μM de DOX por diferentes períodos (0, 0,5, 2, 6, 12, 24 e 48 h) para detecção da expressão de RNA.…”