LncRNA DANCR reduces anoxia-induced rat myocardial cell line H9c2 injury by targeting miR-3646

doi:10.16352/j.issn.1001-6325.2023.01.0144

Abstract

Abstract: Objective To study the role of long non-coding RNA (lncRNA) differentiation antagonizing non-coding RNA (DANCR) in myocardial ischemia and the targeted regulation of microRNA-3646 (miR-3646). Methods H9c2 cells(rat cardiomyocytes cell line)were separated into 6 groups: control group, anoxia group, pc-DANCR (DANCR over-expression) group, pc-NC (DANCR over-expression control) group, miR-3646 mimic (miR-3646 mimicry) group, mimic NC (miR-3646 mimic control) group, and pc-DANCR+miR-3646 mimic group; RT-qPCR method was implemented to measure the expression of lncRNA DANCR and miR-3646 to determine the transfection effect; Flow cytometry was performed to measure the rate of apoptosis; CCK-8 assay was implemented to cell measure survival; Electron microscopy was applied to observe the structural injury of H9c2 cells; Western blot was performed to measure the expression of activated cleaved caspase-3 (cleaved caspase-3) and heme oxygenase-1 (Ho-1). Dual-luciferase experiments were performed to verify the targeting relationship of miR-3646 with lncRNA DANCR and Ho-1. RNA interference Ho-1 (ShHo-1) and pc-DANCR were co-transfected into H9c2 cells, and the cell viability was measured. Results Compared with the control group, the injury and apoptosis of H9c2 cells in the anoxia group were serious, the survival rate and the expression of lncRNA DANCR were decreased, and the expression of miR-3646 was increased (P<0.05). Compared with the anoxia group, the injury and apoptosis of H9c2 cells in the pc-DANCR group were alleviated, the survival rate was increased, the expression of anti-injury protective factor Ho-1 was increased, and the expression of miR-3646 was decreased(P<0.05). The injury and apoptosis of H9c2 cells in the miR-3646 mimic group were further aggravated, and miR-3646 mimic was able to attenuate the anti-injury and anti-apoptotic effects of pc-DANCR (P<0.05). There was a targeted regulatory relationship between lncRNA DANCR, Ho-1 and miR-3646. Interfering with Ho-1 attenuated the promoting effect of lncRNA-DANCR on the survival of H9c2 cells induced by anoxia (P<0.05). Conclusions LncRNA DANCR can down-regulate the expression of miR-3646 and promote the expression of Ho-1 to alleviate anoxia-induced H9c2 cells injury and improve cell survival.

Key words: long non-coding RNA, differentiation antagonist non-coding RNA, microRNA-3646, cardiomyocytes, anoxia induction

CLC Number:

R541

QIU Libin, ZHANG Xincai, ZHAO Qian. LncRNA DANCR reduces anoxia-induced rat myocardial cell line H9c2 injury by targeting miR-3646[J]. Basic & Clinical Medicine, 2023, 43(1): 144-151.

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