Expression Levels of hsa-miR-590-3p in Hepatocellular Carcinoma and Osteosarcoma Cell Lines and its Downstream Target Genes


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Mennatallah Elfar1,Asma Amleh2
1Generations Labs,2The American University in Cairo

Abstract

Background

Small non-coding RNA sequences, known as mircoRNAs (miRNAs), of around 18-25 nucleotides can regulate various cellular pathways by acting on tumor suppressors, oncogenes or both. miRNAs are generally tissue-specific and can be up-regulated or down-regulated, depending on the cancer or the tissue in which they are found. In this study, we focused on hsa-miR-590-3p. hsa-miR-590-3p was found to be involved in several types of cancers and controls several downstream target genes.

Method

Several bioinformatics tools were used to identify potential downstream target genes of hsa-miR-590-3p. Using miRanda-mirSVR, Expression Atlas and The Human Protein Atlas, the following genes (SOX2, N-cadherin, E-cadherin and FOXA2) were utilized as potential downstream target genes of hsa-miR-590-3p. Various molecular techniques were carried out to further validate our in silico results using SNU449 and HepG2, hepatocellular carcinoma cell lines, and U2OS, an osteosarcoma cell line. RT-PCR and western blotting techniques were used to detect the mRNA and protein expression levels of these genes respectively. Co-localization of hsa-miR-590-3p and its potential downstream target gene, SOX2, was conducted using miRNA in situ hybridization combined with immunohistochemistry staining using anti-SOX2.

Results

An inverse correlation between hsa-miR-590-3p expression and SOX2 protein expression in SNU449 and U2OS was observed.

Conclusion

Studying the expression of hsa-miR-590-3p downstream target genes can increase our understanding of cancer pathogenesis and how it can be utilized as a therapeutic tool.