Abstract

N⁶-methyladenosine (m⁶A), a methylation in the N⁶ position of adenosine especially in the mRNA, exerts diverse physiological and pathological functions. However, the precise role of m⁶A methylation in hypoxic preconditioning (HPC) is still unknown. Here, we observed that HPC treatment protected H9c2 cells against H₂O₂-induced injury, upregulated the m⁶A level in the total RNA and the expression of methyltransferase like 3 (METTL3), methyltransferase like 14 (METTL14), and long noncoding RNA (lncRNA) H19. Either knockdown of METTL3 or METTL14 notably reversed the HPC-induced enhancement of cell viability, anti-apoptosis ability, and H19 expression. Methylated RNA immunoprecipitation (IP) indicated that knockdown of METTL3 or METTL14 decreased m⁶A level in the lncRNA H19. Gain-of-function assay demonstrated that H19 overexpression could partially rescue the decreased protection mediated by METTL3 or METTL14 knockdown in HPC-treated H9c2 cells. RNA binding protein immunoprecipitation (RIP) assay showed that METTL3 and METTL14 could directly bind with H19. Our study identified a novel pattern of posttranscriptional regulation in HPC treatment. Since METTL3, METTL14, and lncRNA H19 were involved in HPC protection, they could be considered as potential biomarkers and therapeutic targets in HPC-derived cardiac rehabilitation and therapeutic approaches.

中文翻译：

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N6-甲基腺苷甲基转移酶部分通过与lncRNA H19的相互作用在缺氧预处理中起作用

摘要

N ⁶ -甲基腺苷(m ⁶ A)是腺苷N ⁶位置的甲基化，尤其是在 mRNA 中，具有多种生理和病理功能。然而，m ⁶ A 甲基化在缺氧预处理 (HPC) 中的确切作用仍然未知。在这里，我们观察到 HPC 处理保护 H9c2 细胞免受 H ₂ O ₂诱导的损伤，上调 m ⁶总 RNA 水平和甲基转移酶样 3 (METTL3)、甲基转移酶样 14 (METTL14) 和长链非编码 RNA (lncRNA) H19 的表达。METTL3 或 METTL14 的敲低显着逆转了 HPC 诱导的细胞活力、抗细胞凋亡能力和 H19 表达的增强。甲基化 RNA 免疫沉淀 (IP) 表明 METTL3 或 METTL14 的敲低降低了 m ⁶lncRNA H19 中的一个水平。功能获得试验表明，H19 过表达可以部分挽救由 HPC 处理的 H9c2 细胞中 METTL3 或 METTL14 敲低介导的保护降低。RNA结合蛋白免疫沉淀（RIP）分析表明METTL3和METTL14可以直接与H19结合。我们的研究确定了 HPC 治疗中转录后调控的新模式。由于 METTL3、METTL14 和 lncRNA H19 参与 HPC 保护，因此它们可以被视为 HPC 衍生的心脏康复和治疗方法中潜在的生物标志物和治疗靶点。