MiR-612, miR-637, and miR-874 can Regulate VEGFA Expression in Hepatocellular Carcinoma Cell Lines.

Márcia Maria U. Castanhole-Nunes,Marlon F. Mattos,André R. C. P. Oliveira,Renato F. da Silva,Ana Lívia S. Galbiatti-Dias,Nathalia M. Tunissiolli,Eny M. Goloni-Bertollo,Erika C. Pavarino,Rosa S. Kawasaki-Oyama

doi:10.3390/genes13020282

Márcia Maria U. Castanhole-Nunes, Marlon F. Mattos + Show 7 more

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https://doi.org/10.3390/genes13020282

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Abstract

MicroRNAs (miRNAs) are short non-coding RNA molecules acting as important posttranscriptional gene and protein expression regulators in cancer. The study goal was to examine VEGFA (vascular endothelial growth factor A) expression in hepatocellular carcinoma (HCC) cell lines upon transfection miR-612, miR-637, or miR-874. Methods: MiR-612 mimics, miR-637 mimics, or miR-874 inhibitors were transfected using Lipofectamine RNAiMax in both HCC cell lines, HepG2 and HuH-7. Real-time PCR, Western blotting, and ELISA methods were used to evaluate VEGFA regulation by the miRNAs. Results: Gene and protein expression levels of VEGFA were down-expressed in both cell lines, HepG2 and HuH-7, transfected with miR-612 or miR-637. Transfection with miR-874 inhibitor showed an increase in VEGFA gene expression in HepG2 and HuH-7 cell lines; however, no regulation was observed on VEGFA protein expression by miR-874 inhibition. Correlation analysis between miRNAs and VEGFA protein expression showed that miR-637 and miR-874 expression present inversely correlated to VEGFA protein expression. Conclusions: VEGFA was down-regulated in response to hsa-miR-612 or hsa-miR-637 overexpression; however, the modulation of VEGFA by miR-874 was observed only at the gene expression and thus, needs further investigation.

Highlights

Publisher’s Note: MDPI stays neutralHepatocellular carcinoma (HCC) is the most common primary liver malignancy and is a leading cause of cancer-related deaths worldwide [1]
Because the inhibition of vascular endothelial growth factor A (VEGFA) signaling interferes in the angiogenesis process, and miRNAs may provide a potential anti-angiogenesis therapy for cancer treatment, we evaluated the expression of VEGFA in hepatocellular carcinoma (HCC) cell lines upon treatment with miR-612 and miR-637 mimics and miR-874 inhibitors
The analysis of The Cancer Genome Atlas Program (TCGA) database through the UALCAN [22] website showed that there is a positive correlation between KRAS (Figure 1a), AKT1 (Figure 1b), and STAT3 (Figure 1c) genes in Liver Hepatocellular carcinoma (LIHC)

Summary

Introduction

Hepatocellular carcinoma (HCC) is the most common primary liver malignancy and is a leading cause of cancer-related deaths worldwide [1]. MicroRNAs (miRNAs) are short non-coding RNA molecules acting as important posttranscriptional gene and protein expression regulators [4,5,6,7]. Some miRNAs can regulate the vascular endothelial growth factor A (VEGFA) gene, which acts in blood vessel growth in some cancer types, including hepatocellular carcinoma [8,9,10,11,12]. Studies in cancer, including HCC, have shown that miRNAs have an essential role in angiogenesis, tumorigenesis, and metastasis [13,14,15]. A review highlighted that miRNAs are meaningful for regulating particular endothelial processes downstream of VEGF, representing with regard to jurisdictional claims in published maps and institutional affiliations

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