Up-regulation of miR-381 inhibits NAD+ salvage pathway and promotes apoptosis in breast cancer cells

Authors

  • Zahra Bolandghamat Pour Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran
  • Mitra Nourbakhsh Department of Biochemistry, Faculty of Medicine, Iran University of Medical Sciences, Hemmat Highway 1449614535, Tehran, Iran, Tel: +98 21 86703109, Fax: +98 21 88622742, Mobile: +98 912 2874740, E-mail: nourbakhsh.m@iums.ac.ir
  • Kazem Mousavidzadeh Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran; Cellular and Molecular Research Center, Faculty of Medicine, Iran University of Medical Sciences ,Tehran, Iran. Hemmat Highway 1449614535, Tehran, Iran, Tel: +98-21-86704720, Fax: +98-21-88622578, Mobile: +98-9369973054, E-mail: mousavizadeh.k@iums.ac.ir
  • Zahra Madjd Department of Molecular Medicine, Faculty of Advanced Technologies in Medicine, Iran University of Medical Sciences, Tehran, Iran; Cellular and Molecular Research Center, Faculty of Medicine, Iran University of Medical Sciences, Tehran, Iran
  • Seyedeh Sara Ghorbanhosseini Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran
  • Zohreh Abdolvahabi Department of Biochemistry and Genetics, Cellular and Molecular Research Center, Qazvin University of Medical Sciences, Qazvin, Iran
  • Zahra Hesari Laboratory Sciences Research Center, Golestan University of Medical Sciences, Gorgan, Iran; Department of Laboratory Sciences, Faculty of Paramedicine, Golestan University of Medical Sciences, Gorgan, Iran
  • Samira Ezzati Mobaser Department of Biochemistry, School of Medicine, Iran University of Medical Sciences, Tehran, Iran

DOI:

https://doi.org/10.17179/excli2019-1431

Keywords:

Nicotinamide phosphoribosyltransferase, nicotinamide adenine dinucleotide, miR-381, apoptosis, breast cancer, tumor suppression

Abstract

Nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme involved in nicotinamide adenine dinucleotide (NAD) salvage pathway, is overexpressed in many human malignancies such as breast cancer. This enzyme plays a critical role in survival and growth of cancer cells. MicroRNAs (miRNAs) are among the most important regulators of gene expression, and serve as potential targets for diagnosis, prognosis, and therapy of breast cancer. Therefore, the aim of this study was to assess the effect of NAMPT inhibition by miR-381 on breast cancer cell survival. MCF-7 and MDA-MB-231 cancer cell lines were transfected with miR-381 mimic, inhibitor, and their corresponding negative controls (NCs). Subsequently, the level of NAMPT and NAD was assessed using real-time PCR, immuno-blotting, and enzymatic methods, respectively. In order to evaluate apoptosis, cells were labelled with Annexin V-FITC and propidium iodide and analyzed by flow cytometry. Bioinformatics analysis was performed to recognize whether NAMPT 3′-untranslated region (UTR) is a direct target of miR-381 and the results were authenticated by the luciferase reporter assay using a vector containing the 3′-UTR sequence of NAMPT. Our results revealed that the 3′-UTR of NAMPT was a direct target of miR-381 and its up-regulation decreased NAMPT gene and protein expression, leading to a notable reduction in intracellular NAD and subsequently cell survival and induction of apoptosis. It can be concluded that miR-381 has a vital role in tumor suppression by down-regulation of NAMPT, and it can be a promising candidate for breast cancer therapy.

Published

2019-08-27

How to Cite

Bolandghamat Pour, Z., Nourbakhsh, M., Mousavidzadeh, K., Madjd, Z., Ghorbanhosseini, S. S., Abdolvahabi, Z., Hesari, Z., & Mobaser, S. E. (2019). Up-regulation of miR-381 inhibits NAD+ salvage pathway and promotes apoptosis in breast cancer cells. EXCLI Journal, 18, 683–696. https://doi.org/10.17179/excli2019-1431

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Original articles

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