Department of General Practice Medicine, The Third Hospital of Hebei Medical University, 050051, Shijiazhuang, China.
Department of Cardiology, The Third Hospital of Hebei Medical University, 050051, Shijiazhuang, China.
Lab Invest. 2022 Aug;102(8):782-793. doi: 10.1038/s41374-022-00740-8. Epub 2022 Feb 11.
DNA methyltransferase 1 (DNMT1) shows close link with heart disease. This study aimed to define the role DNMT1 plays in heart failure and determine the underlying mechanism. Expression of microRNA (miR)-152-3p, DNMT1, E26 transformation specific-1 (ETS1) and ras homolog gene family member H (RhoH) was determined by RT-qPCR and/or western blot analysis. The interaction between miR-152-3p and ETS1 was predicted and verified. Methylation of the miR-152-3p promoter region was assessed using methylation-specific PCR. H9c2 cells were chosen for in vitro assays to examine the regulatory role of DNMT1 in autophagy and mitophagy with respect to miR-152-3p/ETS1/RhoH. Doxorubicin (DOX)-induced rat models of heart failure were employed for in vivo validation. DNMT1 expression was upregulated in the heart tissues of DOX-induced rats, where it showed an inverse correlation with miR-152-3p expression. Moreover, DNMT1 was shown to enhance methylation of the miR-152-3p promoter region and suppress its expression, leading to inhibition of mitophagy in H9c2 cells. In addition, DNMT1 enhanced expression of ETS1, which further elevated RhoH expression. Moreover, ETS1-elevated RhoH reduced cell viability and promoted autophagy and mitophagy in H9c2 cells upon treatment with DOX. Next, in vivo results demonstrated that depletion of DNMT1 protected rats from heart failure in a miR-152-3p/ETS1/RhoH-dependent manner. Overall, these findings indicate that DNMT1 may inhibit expression of miR-152-3p by promoting the methylation of miR-152-3p and enhancing the expression of ETS1, thereby inducing RHOH transcriptional activation and inhibiting mitochondrial autophagy, ultimately promoting the development of heart failure.
DNA 甲基转移酶 1（DNMT1）与心脏病密切相关。本研究旨在确定 DNMT1 在心力衰竭中的作用，并确定其潜在机制。通过 RT-qPCR 和/或 Western blot 分析测定 microRNA（miR）-152-3p、DNMT1、E26 转化特异性 1（ETS1）和 ras 同源基因家族成员 H（RhoH）的表达。预测和验证了 miR-152-3p 与 ETS1 之间的相互作用。使用甲基化特异性 PCR 评估 miR-152-3p 启动子区域的甲基化。选择 H9c2 细胞进行体外测定，以检查 DNMT1 在自噬和线粒体自噬方面对 miR-152-3p/ETS1/RhoH 的调节作用。使用阿霉素（DOX）诱导的心力衰竭大鼠模型进行体内验证。在 DOX 诱导的大鼠心脏组织中上调了 DNMT1 的表达，其与 miR-152-3p 的表达呈负相关。此外，DNMT1 被证明增强了 miR-152-3p 启动子区域的甲基化并抑制其表达，从而抑制 H9c2 细胞中的线粒体自噬。此外，DNMT1 增强了 ETS1 的表达，进一步提高了 RhoH 的表达。此外，ETS1 上调的 RhoH 降低了细胞活力，并在 H9c2 细胞用 DOX 处理后促进自噬和线粒体自噬。接下来，体内结果表明，DNMT1 的耗竭以 miR-152-3p/ETS1/RhoH 依赖的方式保护大鼠免受心力衰竭。总的来说，这些发现表明，DNMT1 可能通过促进 miR-152-3p 的甲基化和增强 ETS1 的表达来抑制 miR-152-3p 的表达，从而诱导 RHOH 转录激活并抑制线粒体自噬，最终促进心力衰竭的发展。
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