miRNAs are small non-coding RNA sequences of 18-25 nucleotides. They can regulate different cellular pathways by acting on tumor suppressors, oncogenes, or both. miRNAs are mostly tissue-specific, and their expression varies depending on the cancer or the tissue in which they are found. hsa-miR-590-3p was found to be involved in several types of cancers. In this study, we identified potential downstream target genes of hsa-miR-590-3p computationally. Several bioinformatics tools and more than one approach were used to identify potential downstream target genes of hsa-miR-590-3p. CX3CL1, SOX2, N-cadherin, E-cadherin, and FOXA2 were utilized as potential downstream target genes of hsa-miR-590-3p. SNU449 and HepG2, hepatocellular carcinoma cell lines, were used to carry out various molecular techniques to further validate our in silico results. mRNA and protein expression levels of these genes were detected using RT-PCR and western blotting, respectively. Co-localization of hsa-miR-590-3p and its candidate downstream target gene, SOX2, was carried out using a miRNA in situ hybridization combined with immunohistochemistry staining through anti-SOX2. The results show that there is an inverse correlation between hsa-miR-590-3p expression and SOX2 protein expression in SNU449. Subsequently, we suggest that SOX2 can be a direct downstream target of has-miR-590-3p indicating that it may have a role in the self-renewal and self-maintenance of cancer cells. We also suggest that CX3CL1, E-cadherin, N-cadherin, and FOXA2 show a lot of potential as downstream target genes of hsa-miR-590-3p signifying its role in epithelial-mesenchymal transition. Studying the expression of hsa-miR-590-3p downstream targets can enrich our understanding of the cancer pathogenesis and how it can be used as a therapeutic tool.

中文翻译：

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HCC 细胞系中的 miR-590-3p 及其下游靶基因。

miRNA 是 18-25 个核苷酸的小非编码 RNA 序列。它们可以通过作用于肿瘤抑制基因、癌基因或两者来调节不同的细胞途径。miRNA 大多数是组织特异性的，它们的表达根据癌症或它们所在的组织而变化。hsa-miR-590-3p被发现与多种类型的癌症有关。在本研究中，我们通过计算确定了 hsa-miR-590-3p 的潜在下游靶基因。使用多种生物信息学工具和不止一种方法来识别 hsa-miR-590-3p 的潜在下游靶基因。CX3CL1、SOX2、N-钙粘蛋白、E-钙粘蛋白和 FOXA2 被用作 hsa-miR-590-3p 的潜在下游靶基因。SNU449 和 HepG2（肝细胞癌细胞系）被用来进行各种分子技术，以进一步验证我们的计算机结果。分别使用RT-PCR和蛋白质印迹法检测这些基因的mRNA和蛋白质表达水平。使用 miRNA 原位杂交结合抗 SOX2 免疫组织化学染色，对 hsa-miR-590-3p 及其候选下游靶基因 SOX2 进行共定位。结果表明，SNU449中hsa-miR-590-3p表达量与SOX2蛋白表达量呈负相关。随后，我们认为SOX2可能是has-miR-590-3p的直接下游靶标，表明它可能在癌细胞的自我更新和自我维持中发挥作用。我们还认为 CX3CL1、E-cadherin、N-cadherin 和 FOXA2 作为 hsa-miR-590-3p 下游靶基因显示出很大的潜力，表明其在上皮间质转化中的作用。研究 hsa-miR-590-3p 下游靶标的表达可以丰富我们对癌症发病机制及其如何用作治疗工具的理解。