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Methotrexate-induced apoptosis in human ovarian adenocarcinoma SKOV-3 cells via ROS-mediated bax/bcl-2-cyt-c release cascading

Authors AlBasher G, AlKahtane AA, Alarifi S, Ali D, Alessia MS, Almeer RS, Abdel-Daim MM, Al-Sultan NK, Al-Qahtani AA, Ali H, Alkahtani S

Received 27 June 2018

Accepted for publication 3 October 2018

Published 17 December 2018 Volume 2019:12 Pages 21—30

DOI https://doi.org/10.2147/OTT.S178510

Checked for plagiarism Yes

Review by Single-blind

Peer reviewers approved by Dr Justinn Cochran

Peer reviewer comments 1

Editor who approved publication: Dr Takuya Aoki


Gadah AlBasher,1 Abdullah A AlKahtane,1 Saud Alarifi,1 Daoud Ali,1 Mohammed S Alessia,2 Rafa S Almeer,1 Mohamed M Abdel-Daim,3 Nouf K Al-Sultan,1 Ahmed A Al-Qahtani,4,5 Huma Ali,6 Saad Alkahtani1

1Department of Zoology, College of Science, King Saud University, Riyadh, Saudi Arabia; 2Department of Biology, Science College, Al-Imam Muhammad Ibn Saud, Islamic University, Riyadh, Saudi Arabia; 3Department of Pharmacology, Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Egypt; 4Department of Infection and Immunity, Research Center, King Faisal Specialist Hospital and Research Center, Riyadh, Saudi Arabia; 5Department of Microbiology and Immunology, Alfaisal University School of Medicine, Riyadh, Saudi Arabia; 6Department of Chemistry Maulana Azad National Institute of Technology, Bhopal, India

Introduction: The communication between a substance and a cell may depend on whether the cell is normal or pathological. The disease cells and drug interaction may occasionally overcome beneficial action of the drug; subsequently, it is important to investigate the effect of the drug in both the normal and target cells. This study aimed to evaluate the methotrexate (MTX) antiproliferative effect and explore the mechanistic approach to investigate the cell death index in SKOV-3 ovarian cells during treatment with MTX.
Methods:
In vitro studies of SKOV-3 cells were examined by tetrazolium assay after exposure to various concentrations of MTX. Moreover, reactive oxygen species (ROS) generation, mitochondrial membrane potential, DNA damage, and AO/EtBr staining morphological analysis of necrotic/apoptotic cells were detected; cellular impairment in mitochondria and DNA was confirmed by JC-1 mitotracker/DAPI, respectively, and cell death pathway markers; bax/bcl-2 were analyzed.
Results: A dose-dependent antiproliferative effect of MTX treatment was observed in SKOV-3 cells; the prominent inhibitory concentration was 40 µM of MTX (P<0.01). The growth inhibition rates of the cancer cells reached 24.07% in MTX. The MTX showed increase in ROS generation and mitochondrial depolarization, and DNA integrity cells collectively advocated the apoptotic cell death at higher concentration. In addition, the results of reverse transcription polymerase chain reaction also supported the apoptosis by upregulating the bax and downregulating the bcl-2 (P<0.01). Thus the MTX moderately provokes apoptosis.
Conclusion: Our findings suggest that MTX acts on SKOV-3 cancer cells by increasing intracellular ROS levels, leading to DNA damage and altering the MMP along with apoptotic gene upregulation. This mechanism may provide new therapeutic targets to improve tumor treatment.

Keywords: methotrexate, MMP, apoptosis, ROS, SKOV-3 cells

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