Down-regulation of NAMPT expression by mir-206 reduces cell survival of breast cancer cells

Hesari, Z. and Nourbakhsh, M. and Hosseinkhani, S. and Abdolvahabi, Z. and Alipour, M. and Tavakoli-Yaraki, M. and Ghorbanhosseini, S.S. and Yousefi, Z. and Jafarzadeh, M. and Yarahmadi, S. (2018) Down-regulation of NAMPT expression by mir-206 reduces cell survival of breast cancer cells. Gene, 673. pp. 149-158.

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Nicotinamide adenine dinucleotide (NAD) is a critical coenzyme for all living cells. Nicotinamide phosphoribosyltransferase (NAMPT) functions as a key enzyme in the salvage pathway of NAD biosynthesis. Cancer cells have higher rate of NAD consumption and therefore NAMPT is essential for their survival. Thus, we investigated the effect of NAMPT inhibition by miR-206 on breast cancer cell survival. Breast cancer cells were transfected with miR-206 mimic, inhibitor and their negative controls. NAMPT levels were assessed by real-time PCR as well as western blotting. Cell survival assay and quantification of NAD level were performed by using colorimetric methods. Apoptosis assay was performed by labeling cells with Annexin V-FITC and propidium iodide followed by the flow cytometric analysis. Bioinformatics analysis was done to assess whether NAMPT 3�-UTR is a direct target of miR-206 and the results were confirmed by the luciferase reporter assay. NAMPT 3�-UTR was shown to be a direct target of miR-206. miR-206 reduced NAMPT expression at the protein level, leading to a significant decrease in the intracellular NAD level and subsequent decline in cell survival and induction of apoptosis. Targeting of NAMPT-mediated NAD salvage pathway by miR-206 might provide a new insight in the possible molecular mechanism of breast cancer cell growth regulation. This pathway might provide a new approach for breast cancer therapy. © 2018

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: fluorescein isothiocyanate; lipocortin 5; messenger RNA; microRNA; microRNA 206; nicotinamide phosphoribosyltransferase; propidium iodide; unclassified drug; cytokine; luciferase; microRNA; MIRN206 microRNA, human; nicotinamide phosphoribosyltransferase; nicotinamide phosphoribosyltransferase, human, 3' untranslated region; apoptosis assay; Article; bioinformatics; breast cancer cell line; cell labeling; cell survival; cell viability; colorimetry; controlled study; down regulation; enzyme inhibition; flow cytometry; gene expression regulation; luciferase assay; MCF-10A cell line; MCF-7 cell line; MDA-MB-231 cell line; NAMPT gene; priority journal; protein expression level; real time polymerase chain reaction; translation regulation; upregulation; Western blotting; apoptosis; breast tumor; cell survival; female; gene expression profiling; genetics; HEK293 cell line; human; metabolism; mortality; tumor cell line, 3' Untranslated Regions; Apoptosis; Blotting, Western; Breast Neoplasms; Cell Line, Tumor; Cell Survival; Cytokines; Female; Gene Expression Profiling; Gene Expression Regulation, Neoplastic; HEK293 Cells; Humans; Luciferases; MCF-7 Cells; MicroRNAs; Nicotinamide Phosphoribosyltransferase
Subjects: WP Gynecology
Depositing User: eprints admin
Date Deposited: 25 Dec 2018 13:26
Last Modified: 19 Aug 2019 10:56

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