A novel mechanism underlying alcohol dehydrogenase expression: hsa-miR-148a-3p promotes ADH4 expression via an AGO1-dependent manner in control and ethanol-exposed hepatic cells
A novel mechanism underlying alcohol dehydrogenase expression: hsa-miR-148a-3p promotes ADH4 expression via an AGO1-dependent manner in control and ethanol-exposed hepatic cells
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乙醇脱氢酶表达的新机制：hsa-miR-148a-3p 在对照和乙醇暴露的肝细胞中通过 AGO1 依赖性方式促进 ADH4 表达
DOI：
10.1016/j.bcp.2021.114458
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发表时间：
2021-06-04
影响因子：
5.8
通讯作者：
Yu, Dianke
中科院分区：
文献类型：
--
作者：
Luo, Jiao;Hou, Yufei;Yu, Dianke
乙醇脱氢酶（ADH）和乙醛脱氢酶（ALDHs）在酒精中毒的发生和酒精毒理学中起着重要作用;然而，很少有研究关注酒精代谢酶表达的miRNA介导的机制。在本研究中，我们显示了酒精性肝炎（AH）患者肝脏样品中乙醇脱氢酶（ADH）和乙醛脱氢酶（ALDH）的表达变化，并通过系统的计算机分析预测靶向酒精代谢失调基因的miRNA。预测13种miRNAs分别调节ADH 1A、ADH 4和ALDH 2的表达，其中hsa-miR-148 a-3 p（miR-148 a）在AH患者中表现出最显著的下调。随后使用HepG 2细胞的实验证据证明，miR-148 a通过直接结合ADH 4的编码序列并通过AGO 1依赖性方式增加mRNA的稳定性来促进ADH 4的表达。进一步的分析表明，ADH 4转录物的二级结构影响miR-148 a-3 p的靶可及性和结合。综上所述，我们的研究结果表明AH患者酒精代谢关键酶的表达受到抑制，miR 148 a对ADH 4的非经典正调控揭示了ADH基因的一种新的调控机制。
The alcohol dehydrogenases (ADHs) and aldehyde dehydrogenases (ALDHs) play critical roles in alcoholism development and alcohol toxicology; however, few studies have focused on the miRNA-mediated mechanisms underlying the expressions of alcohol-metabolizing enzymes. In the present study, we showed the expression changes of each alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) in the liver samples of alcoholic hepatitis (AH) patients, and predicted the miRNAs targeting the dysregulated alcohol-metabolizing genes by a systematic in silico analysis. 13 miRNAs were predicted to regulate the expressions of ADH1A, ADH4, and ALDH2, respectively, with hsa-miR-148a-3p (miR-148a) showing the most significant downregulation in AH patients. Following experimental evidence using HepG2 cells proved that miR-148a promoted ADH4 expression by directly binding to the coding sequence of ADH4 and increasing the mRNA stability via an AGO1-dependent manner. Additional assays showed that secondary structure of ADH4 transcript affected the target accessibility and binding of miR-148a-3p. In sum, our results suggest that the expressions of key alcohol-metabolizing enzymes are repressed in AH patients, and the non-canonical positive regulation of miR148a on ADH4 reveals a new regulationary mechanism for ADH genes.