DCC netrin 1 receptor (DCC) affects the structure and function of the dopamine circuitry, which in turn affects the susceptibility to developing addiction. In a previous study, we found that single nucleotide polymorphism (SNP) rs12607853 in the 3′ untranslated region (3′-UTR) of DCC was significantly associated with heroin addiction. In the current study, we first used bioinformatics prediction to identify the DCC rs12607853 C allele as a potential hsa-miR-422a and hsa-miR-378c target site. We then used vector construction and dual-luciferase reporter assays to investigate the targeting relationship of DCC rs12607853 with hsa-miR-422a and hsa-miR-378c. The dual-luciferase reporter gene assay confirmed that the C allele of rs12607853 in combination with hsa-miR-422a led to repressed dual-luciferase gene expression. Moreover, gene expression assays disclosed that hsa-miR-422a inhibited DCC expression at both the mRNA and protein levels. We also found that morphine inhibited the expression of hsa-miR-422a but increased the expression of DCC mRNA, and this change in the expression of hsa-miR-422a could not be reversed by naloxone, which suggested that the role of DCC in opioid addiction might be regulated by hsa-miR-422a. In summary, this study improves our understanding of the role of hsa-miR-422a and identifies the genetic basis of rs12607853, which might contribute to the discovery of new biomarkers or therapeutic targets for opioid addiction.

中文翻译：

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microRNA-422a以依赖于 SNP 的方式抑制DCC表达 rs12607853

DCC netrin 1 受体 (DCC) 影响多巴胺回路的结构和功能，进而影响成瘾的易感性。在之前的一项研究中，我们发现 DCC 3' 非翻译区 (3'-UTR) 中的单核苷酸多态性 (SNP) rs12607853 与海洛因成瘾显着相关。在目前的研究中，我们首先使用生物信息学预测将 DCC rs12607853 C 等位基因鉴定为潜在的 hsa-miR-422a 和 hsa-miR-378c 靶位点。然后我们使用载体构建和双荧光素酶报告基因检测来研究 DCC rs12607853 与 hsa-miR-422a 和 hsa-miR-378c 的靶向关系。双荧光素酶报告基因检测证实，rs12607853 的 C 等位基因与 hsa-miR-422a 结合导致双荧光素酶基因表达受到抑制。而且，基因表达分析表明 hsa-miR-422a 在 mRNA 和蛋白质水平上抑制 DCC 表达。我们还发现吗啡抑制了 hsa-miR-422a 的表达，但增加了 DCC mRNA 的表达，而这种 hsa-miR-422a 表达的变化不能被纳洛酮逆转，这表明 DCC 在阿片类药物中的作用成瘾可能受 hsa-miR-422a 调节。总之，这项研究提高了我们对 hsa-miR-422a 作用的理解，并确定了 rs12607853 的遗传基础，这可能有助于发现新的生物标志物或阿片类药物成瘾的治疗靶点。并且 hsa-miR-422a 表达的这种变化不能被纳洛酮逆转，这表明 DCC 在阿片类药物成瘾中的作用可能受 hsa-miR-422a 的调节。总之，这项研究提高了我们对 hsa-miR-422a 作用的理解，并确定了 rs12607853 的遗传基础，这可能有助于发现新的生物标志物或阿片类药物成瘾的治疗靶点。hsa-miR-422a 表达的这种变化不能被纳洛酮逆转，这表明 DCC 在阿片类药物成瘾中的作用可能受 hsa-miR-422a 的调节。总之，这项研究提高了我们对 hsa-miR-422a 作用的理解，并确定了 rs12607853 的遗传基础，这可能有助于发现新的生物标志物或阿片类药物成瘾的治疗靶点。