ALTERING THE EXPRESSION OF P-GLYCOPROTEIN RESPONSIBLE FOR CHEMOTHERAPEUTIC DRUG RESISTANCE IN HEPG2 CELL LINE POST-TREATMENT WITH UREA AND β-MERCAPTOETHANOL
Keywords:HepG2 cells,, P-glycoprotein pumps,, Protein expression,, Nil
Objective: The objective of this study is to alter the expression of p-glycoprotein (p-gp) pump proteins in HepG2 cells after treating with urea and β-mercaptoethanol (BME) (lead compounds). The most common cause for resistance to a broad range of anticancer drugs is influenced by overexpression of p-gp pumps that detect and eject anticancer drugs from the cancer cell. Altering the expression of these proteins will reduce the efflux action and enhance the drug retention eventually killing the cancer cell.
Materials and Methods: 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyl tetrazolium bromide (MTT) assay was carried out to measure the cell viability (HepG2 cells) post-treatment with the lead compounds followed by flow cytometric analysis for protein expression studies.
Results: MTT assay confirms that the viability of HepG2 cells reduces as the concentrations of the lead compounds are increased. Flow cytometric analysis confirms reduced p-gp expression in HepG2 cells post-treatment with urea and BME. Compare to BME, urea turns out to be a potential compound in altering the expression of p-gp.
Conclusion: The present cell line study confirms that urea and BME are potential compounds which are able to reduce the p-gp expression inHepG2 cells.
Chang G, Roth CB. Structure of MsbA from E. coli: A homolog of the multidrug resistance ATP binding cassette (ABC) transporters. Science 2001;293:1793-800.
Sauna ZE, Ambudkar SV. Characterization of the catalytic cycle of ATP hydrolysis by human P-glycoprotein. The two ATP hydrolysis events in a single catalytic cycle are kinetically similar but affect different functional outcomes. J Biol Chem 2001;276:11653-61.
Borst P, Elferink RO. Mammalian ABC transporters in health and disease. Annu Rev Biochem 2002;71:537-92.
Schinkel AH, Smit JJ, van Tellingen O, Beijnen JH, Wagenaar E, van Deemter L, et al. Disruption of the mouse mdr1a P-glycoprotein gene leads to a deficiency in the blood-brain barrier and to increased sensitivity to drugs. Cell 1994;77:491-502.
Abolhoda A, Wilson A, Ross H, Danenberg PV, Burt M, Scotto KW. Rapid activation of MDR1 gene expression in human metastaticsarcoma after in vivo exposure to doxorubicin. Clin Cancer Res 1999;5:3352-6.
Gottesman MM, Fojo T, Bates SE. Multidrug resistance in cancer: Role of ATP-dependent transporters. Nat Rev Cancer 2002;2:48-58.
Szakács G, Annereau JP, Lababidi S, Shankavaram U, Arciello A,Bussey KJ, et al. Predicting drug sensitivity and resistance: Profiling ABC transporter genes in cancer cells. Cancer Cell 2004;6:129-37.
Hilgendorf C, Ahlin G, Seithel A, Artursson P, Ungell AL, Karlsson J, et al. Expression of thirty-six drug transporter genes in human intestine, liver, kidney, and organotypic cell lines. Drug Metab Dispos2007;35:1333-40.
Haber M, Smith J, Bordow SB, Flemming C, Cohn SL, London WB,et al. Association of high-level MRP1 expression with poor clinical outcome in a large prospective study of primary neuroblastoma. J Clin Oncol 2006;24:1546-53.
Housman G, Byler S, Heerboth S, Lapinska K, Longacre M, Snyder N,et al. Drug resistance in cancer: An overview. Cancers (Basel) 2014;6:1769-92.
Kumar V, Taya P, Neetu. 3D QSAR studies on pyrrolopyrimidines as selective p-glycoprotein antagonist. Int J Pharm Pharm Sci 2014;6:232-9.
Laksmiani NP, Meiyanto ED, Susidarti RA. Cytotoxic activity of Brazile in isolated from secang (Caesalpinia sappan) against MCF7/DOX cells by inhibition of p-glycoprotein. Int J Pharm Pharm Sci 2017;9:124-30.
Szilagyi A, Kardos J, Osvath S, Barna L, Zavodszky P. Protein Folding:The Protein Folding Problem. Berlin Heidelberg: Springer-Verlag;2007. p. 4-6.
Bowie JU. Membrane proteins: A new method enters the fold. PNAS 2004;101:3995-6.
Kumar KM, Ramakrishnan M, Chakraborty P, Chandramohan V. Docking and dynamic simulation analysis of p-glycoprotein pumpsresponsible for chemotherapeutic resistance post-treatment with urea and β-mercaptoethanol. J Appl Biol Sci 2018;12:26-35.
Alley MC, Scudiere DA, Monks A, Czerwinski M, Shoemaker R, Boyd MR. Validation of an automated microculture tetrazolium assay (MTA) to assess growth and drug sensitivity of human tumor cell lines. Proc Am Assoc Cancer Res 1986;27:389.
Scudiero DA, Shoemaker RH, Paull KD, Monks A, Tierney S, Nofziger TH, et al. Evaluation of a soluble tetrazolium/formazan assay for cell growth and drug sensitivity in culture using human and other tumor cell lines. Cancer Res 1988;48:4827-33.
Shah WA, Dar MD. Antiproliferative and antioxidant activities of Picea smithiana (Wall) Boiss oil. Int J Chem Pharm Sci 2014;2:541-6.
Filipits M, Suchomel RW, Dekan G, Haider K, Valdimarsson G, Depisch D, et al. MRP and MDR1 gene expression in primary breast carcinomas. Clin Cancer Res 1996;2:1231-7.
Beck WT, Grogan TM, Willman CL, Cordon-Cardo C, Parham DM, Kuttesch JF, et al. Methods to detect P-glycoprotein-associated multidrug resistance in patients’ tumors: Consensus recommendations. Cancer Res 1996;56:3010-20.
Chan HS, Haddad G, Zheng L, Bradley G, Dalton WS, Ling V, et al. Sensitive immunofluorescence detection of the expression of P-glycoprotein in malignant cells. Cytometry 1997;29:65-75.
Ishii T, Bannaia S, Sugita Y. Mechanism of growth stimulation of L1210 cells by 2-mercaptoethanol in vitro. J Biol Chem 1981;256:12387-92.
Vogel C, Silva GM, Marcotte EM. Protein expression regulation under oxidative stress. Mol Cell Proteomics 2011;10:M111.009217.
Choudhuri SK. Deactivation of P-glycoprotein by a novel compound, oxalyl bis (N-phenyl) hydroxamic acid. Neoplasma 2002;49:272-7.
Clark PI, Slevin ML, Webb JA, Osborne RJ, Jones S, Wrigley PF, et al. Oral urea in the treatment of secondary tumours in the liver. Br J Cancer 1988;57:317-8.
Click RE. Dietary supplemented 2-mercaptoethanol prevents spontaneous and delays virally-induced murine mammary tumorigenesis. Cancer Biol Ther 2013;14:521-6.
How to Cite
The publication is licensed under CC By and is open access. Copyright is with author and allowed to retain publishing rights without restrictions.