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. 2021 Jul;40(7):1026-1036.
doi: 10.1089/dna.2021.0330. Epub 2021 Jun 8.

MiR-613 Promotes Cell Death in Breast Cancer Cells by Downregulation of Nicotinamide Phosphoribosyltransferase and Reduction of NAD

Shahin Alizadeh-Fanalou  1 , Saman Hosseinkhani  2 , Ali Nazarizadeh  1 , Samira Ezzati-Mobaser  1 , Zahra Hesari  3 , Parisa Aziminezhadan  4 , Zohreh Abdolvahabi  5 , Meysam Abolmaali  6 , Masoumeh Tavakoli-Yaraki  1 , Mitra Nourbakhsh  1   7
Affiliations

MiR-613 Promotes Cell Death in Breast Cancer Cells by Downregulation of Nicotinamide Phosphoribosyltransferase and Reduction of NAD

Shahin Alizadeh-Fanalou et al. DNA Cell Biol. 2021 Jul.

Abstract

NAD is mainly biosynthesized by the enzymatic action of nicotinamide phosphoribosyltransferase (NAMPT) through the salvage pathway. NAD is indispensable for the proper function and metabolism of all living cells, including cancer cells. Our previous researches revealed that inhibition of NAMPT by miRNA (miR) could suppress NAD levels and thereby hinder the growth and promotion of breast cancer (BC). Therefore, the current study was undertaken to investigate the inhibitory effects of miR-613 on NAMPT and BC cells' survival. Bioinformatics analysis and luciferase reporter assay confirmed that NAMPT 3'-untranslated region is a direct target for miR-613. The expression of miR-613 was noticed to be significantly decreased in both clinical tissue samples and BC cells by real-time PCR. Following transfection with miR-613 mimic, the expression of miR-613 was elevated in the BC cells leading to inhibition of NAMPT expression at both mRNA and protein level as measured by real-time PCR and western blotting, respectively. Inhibition of NAMPT led to a remarkable reduction in the concentration of NAD in the BC cells. The transfection also declined cell viability roughly 40% in MD Anderson-Metastatic Breast-231 (MDA-MB-231) cells. Consistently, the apoptosis rate was remarkably increased, around 65% in these cells as assayed by labeling the cells with Annexin V-fluorescein isothiocyanate (FITC) and Propidium Iodide. Targeting the NAMPT-mediated NAD salvage pathway by miR-613 is a novel approach for managing BC, which is worth further investigation.

Keywords: apoptosis; breast cancer; miR-613; miRNA; nicotinamide phosphoribosyltransferase.

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