IN VITRO EVALUATION OF ANTI-CANCER POTENTIAL OF A3 ADENOSINE RECEPTOR AGONIST ON A549 HUMAN LUNG CANCER CELL LINE
DOI:
https://doi.org/10.22159/ijpps.2019v11i6.30863Keywords:
A549 cell line, Adenosine, Doxorubicin, MTT assay, Lung cancerAbstract
Objective: The objective of this study was to evaluate the cytotoxic potential of A3AR agonist (ABMECA) against human lung cancer cell line A549 by using 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay.
Methods: Adenocarcinoma cell line A549 was used to assess MTT based cells viability. In vitro cytotoxic activity was evaluated for 3 different concentration of doxorubicin and A3AR by MTT cytotoxicity assay. Cytotoxicity assay carried out for 3 consecutive days that involves culturing cells into Dulbecco’s MEM medium modified with 10% FBS for 24 h then treatment with different dose of standard and test drug with incubation period of 24 h followed by treatment with MTT for estimation of cytotoxicity and finally, optical density (OD) was measured at 570-630 nm.
Results: Different concentration of doxorubicin (1, 5, 10 µM) and ABMECA (10-6M, 10-5M and 10-4M) shown dose-dependent cytotoxicity. There was a significant decrease (p<0.05) in cell viability in both doxorubicin and ABMECA concentration in a dose-dependent manner. This study may guide further for in vivo evaluation of test drug in the lung cancer model.
Conclusion: A3 Adenosine Receptor agonist could be potential moiety for the treatment of lung cancer and it would require in vivo study for further research.
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References
Ferlay JSI, Ervik M, Dikshit R, Eser S, Mathers C, Rebelo M, et al. Cancer incidence and mortality worldwide: IARC Cancer Base No. 11:GLOBOCAN 2012 v1.0;2012. Available from: http://globocan.iarc.fr. [Last accessed on 21 Jun 2018]
NICPR. Cancer Statistics; 2017. Available from: http://www.cancerindia.org.in/statistics [Last accessed on 21 Jun 2018].
Merighi S, Simioni C, Gessi S, Varani K, Mirandola P, Tabrizi MA, et al. A2B and A3 adenosine receptors modulate vascular endothelial growth factor and interleukin-8 expression in human melanoma cells treated with etoposide and doxorubicin. Neoplasia (New York, NY) 2009;11:1064-73.
Merighi S, Mirandola P, Varani K, Gessi S, Leung E, Baraldi PG, et al. A glance at adenosine receptors: a novel target for antitumor therapy. Pharmacol Ther 2003;100:31-48.
Fishman P, Bar Yehuda S, Synowitz M, Powell JD, Klotz KN, Gessi S, et al. Adenosine receptors and cancer. Hand Exp Pharmacol 2009;193:399-441.
Sacchetto V. The a3 adenosine receptor: a link between inflammation and cancer. Pubblicazionidello IUSS 2010;3:1-149.
Sajjadi FG, Takabayashi K, Foster AC, Domingo RC, Firestein GS. Inhibition of TNF-alpha expression by adenosine: role of A3 adenosine receptors. J Immunol 1996;156:3435-42.
https://blog.quartzy.com/2017/05/01/cell-viability-assays-mtt-protocol. [Last accessed on 05 Apr 2018]
Hwa Yeon Jo. The unreliability of MTT assay in the cytotoxic test of primary cultured glioblastoma cells. Exp Neurobiol 2015;24:235-45.
Ashutosh Bahuguna, Imran Khan, Vivek K Bajpai, Sun Chul Kang. MTT assay to evaluate the cytotoxic potential of a drug. Bangladesh J Pharmacol 2017;12:115-8.
Vistica DT, Skehan P, Scudiero D, Monks A, Pittman A, Boyd MR. Tetrazolium-based assays for cellular viability: a critical examination of selected parameters affecting formazan production. Cancer Res 1991;1:2515–20.
Wang P, Henning SM, Heber D. Limitations of MTT and MTS-based assays for measurement of antiproliferative activity of green tea polyphenols. PLoS One 2010;5:e10202.
Kenneth A Jacobson, Zhan Guo Gao. Adenosine receptors as therapeutic targets. Nat Rev Drug Discovery 2006;5:247–64
Habeela J, Mohan MR. In vitro cytotoxic and apoptosis study of chemical constituents from clerodendrum phlomidis leaf on mcf-7 and a549 cancer cell lines. Asian J Pharm Clin Res 2018;11:299-304.