Effect of Methotrexate and Tea Polyphenols on the Viability and Oxidative Stress in MDA-MB-231 Breast Cancer Cells

Theresa Kelly, Richard Owusu-Apenten

Research output: Contribution to journalArticle

Abstract

Aim: To determine the effect of tea polyphenols and methotrexate on viability and reactive oxygen species (ROS) in a naturally resistant breast cancer cell line MDA-MB-231.Methodology: MDA-MB-231 cells were selected as a model for methotrexate resistant breast cancer. Drug tests were performed over 72 hours at concentrations 0-100 µM. Pre-treatments were with quercetin (QE) or epigallocatechin gallate (EGCG) for 5 hours followed by methotrexate. Cytotoxicity was measured using the MTT assay or resazurin fluorescence assay. ROS was determined using the 2’, 7’-dichlorofluorescein diacetate assay. Intracellular GSH was measured using the monochlorobimane assay.Results: Methotrexate was cytotoxic to MDA-MB-231 cells with IC50 of 35±4 µM. The IC50 value was 68±9.4 µM with QE and 83±16 µM for EGCG. The pre-treatment with QE and EGCG lowered the IC50 for methotrexate by 28% (P =0.009) and 16% (P=0.2027). Intracellular ROS concentrations increased after treatment with methotrexate, QE or EGCG singly and ROS decreased with combination treatment compared with the response for methotrexate only. There were no significant changes in intracellular GSH. Conclusion: Pretreatment with tea polyphenols partially sensitized breast cancer cells towards methotrexate and decreases intracellular ROS. More research is needed to optimize the sensitizing effect of tea phenols on the breast cancer cell response to methotrexate.
LanguageEnglish
Pages152-159
JournalJournal of Applied Life Sciences International
Volume2
Issue number4
Publication statusPublished - 31 Mar 2015

Fingerprint

Polyphenols
Tea
Methotrexate
Oxidative Stress
Breast Neoplasms
Quercetin
Reactive Oxygen Species
Inhibitory Concentration 50
Phenols
Fluorescence
Cell Line
epigallocatechin gallate
Research

Keywords

  • Breast cancer
  • methotrexate
  • quercetin
  • epigallocatechin gallate
  • oxidative stress
  • anticancer effect

Cite this

@article{e8b67ac821a94579aca5f8b88295db27,
title = "Effect of Methotrexate and Tea Polyphenols on the Viability and Oxidative Stress in MDA-MB-231 Breast Cancer Cells",
abstract = "Aim: To determine the effect of tea polyphenols and methotrexate on viability and reactive oxygen species (ROS) in a naturally resistant breast cancer cell line MDA-MB-231.Methodology: MDA-MB-231 cells were selected as a model for methotrexate resistant breast cancer. Drug tests were performed over 72 hours at concentrations 0-100 µM. Pre-treatments were with quercetin (QE) or epigallocatechin gallate (EGCG) for 5 hours followed by methotrexate. Cytotoxicity was measured using the MTT assay or resazurin fluorescence assay. ROS was determined using the 2’, 7’-dichlorofluorescein diacetate assay. Intracellular GSH was measured using the monochlorobimane assay.Results: Methotrexate was cytotoxic to MDA-MB-231 cells with IC50 of 35±4 µM. The IC50 value was 68±9.4 µM with QE and 83±16 µM for EGCG. The pre-treatment with QE and EGCG lowered the IC50 for methotrexate by 28{\%} (P =0.009) and 16{\%} (P=0.2027). Intracellular ROS concentrations increased after treatment with methotrexate, QE or EGCG singly and ROS decreased with combination treatment compared with the response for methotrexate only. There were no significant changes in intracellular GSH. Conclusion: Pretreatment with tea polyphenols partially sensitized breast cancer cells towards methotrexate and decreases intracellular ROS. More research is needed to optimize the sensitizing effect of tea phenols on the breast cancer cell response to methotrexate.",
keywords = "Breast cancer, methotrexate, quercetin, epigallocatechin gallate, oxidative stress, anticancer effect",
author = "Theresa Kelly and Richard Owusu-Apenten",
note = "Reference text: 1. Schmiegelow K. Advances in individual prediction of methotrexate toxicity: a review. Br J Haematol.2009;146(5):489-503. 2. Dadhania VP, Tripathi DN, Vikram A, Ramarao P, Jena GB. Intervention of alpha-lipoic acid ameliorates methotrexate-induced oxidative stress and genotoxicity: A study in rat intestine. Chem Biol Interact. 2010;183(1):85-97. 3. Cakir T, Ozkan E, Dulundu E, Topaloglu U, Sehirli AO, Ercan F, et al. Caffeic acid phenethyl ester (CAPE) prevents methotrexate-induced hepatorenal oxidative injury in rats. J Pharm Pharmacol. 2011;63(12):1566-71. 4. Breast cancer: Incidence, mortality and survival, 2010. Office for National Statistics. 2012; 28:9. Accessed 31 August 2014. Available from: http://www.ons.gov.uk/ons/rel/cancer-unit/breast-cancer-in-england/2010/sum-1.html. 5. Worm J, Kirkin AF, Dzhandzhugazyan KN, Guldberg P. Methylation-dependent silencing of the reduced folate carrier gene in inherently methotrexate-resistant human breast cancer cells. J Biol Chem. 2001;276(43):39990-40000. 6. Thangapazham RL, Singh AK, Sharma A, Warren J, Gaddipati JP, Maheshwari RK. Green tea polyphenols and its constituent epigallocatechin gallate inhibit proliferation of human breast cancer cells in vitro and in vivo. Cancer Lett. 2007;245(1–2):232-41. 7. Stuart EC, Jarvis RM, Rosengren RJ. In vitro mechanism of action for the cytotoxicity elicited by the combination of epigallocatechin gallate and raloxifene in MDA-MB-231 cells. Oncol Rep. 2010;24(3):779-85. 8. Lecumberri E, Deupertuis YM, Mralbell R, Pichard C. Green tea polyphenol epigallocatechin-3-gallate (EGCG) as adjuvant in cancer therapy. Clin Nutr. 2013(32):894-903. 9. Dhanasekaran S. Biswal BK, Sumantran VN, Verma RS. Augmented sensitivity to methotrexate by curcumin induced overexpression of folate receptor in KG-1 cells. Biochimie 2013;95(8): 1567-73. 10. Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods. 1986;89(2):271-7. 11. Magee PJ, Owusu-Apenten R, McCann MJ, Gill CI, Rowland IR. (2012). Chickpea (Cicer arietinum) and other plant derived protease inhibitor concentrates inhibit breast and prostate cancer cell Proliferation in vitro. Nutri Cancer 2012; 64(5): 741-748. 12. Chien SY, Wu YC, Chung JG, Yang JS, Lu HF, Tsou MF, et al. Quercetin-induced apoptosis acts through mitochondrial- and caspase-3-dependent pathways in human breast cancer MDA-MB-231 cells. Hum Exp Toxicol. 2009;28(8):493-503. 13. Murakami A, Ashida H, Terao J. Multi-targeted cancer prevention by quercetin. Cancer Lett. 2008;269(2):315-25. 14. Wu Z, Shah A, Patel N, Yuan X. Development of methotrexate proline prodrug to overcome resistance by MDA-MB-231 cells. Bioorg Med Chem Lett. 2010;20(17):5108-12. 15. Calabrese EJ, Cancer biology and hormesis: Human tumor cell lines commonly display hormetic (biphasic) dose responses. Crit Rev Toxic. 2005;35(6):463-582. 16. Babich H, Schuck AG, Weisburg JH, Zuckerbraun HL. Research strategies in the study of the pro-oxidant nature of polyphenol nutraceuticals. J Toxicol. 2011;2011:467305.",
year = "2015",
month = "3",
day = "31",
language = "English",
volume = "2",
pages = "152--159",
journal = "Journal of Applied Life Sciences International",
issn = "2394-1103",
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}

Effect of Methotrexate and Tea Polyphenols on the Viability and Oxidative Stress in MDA-MB-231 Breast Cancer Cells. / Kelly, Theresa; Owusu-Apenten, Richard.

In: Journal of Applied Life Sciences International, Vol. 2, No. 4, 31.03.2015, p. 152-159.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Effect of Methotrexate and Tea Polyphenols on the Viability and Oxidative Stress in MDA-MB-231 Breast Cancer Cells

AU - Kelly, Theresa

AU - Owusu-Apenten, Richard

N1 - Reference text: 1. Schmiegelow K. Advances in individual prediction of methotrexate toxicity: a review. Br J Haematol.2009;146(5):489-503. 2. Dadhania VP, Tripathi DN, Vikram A, Ramarao P, Jena GB. Intervention of alpha-lipoic acid ameliorates methotrexate-induced oxidative stress and genotoxicity: A study in rat intestine. Chem Biol Interact. 2010;183(1):85-97. 3. Cakir T, Ozkan E, Dulundu E, Topaloglu U, Sehirli AO, Ercan F, et al. Caffeic acid phenethyl ester (CAPE) prevents methotrexate-induced hepatorenal oxidative injury in rats. J Pharm Pharmacol. 2011;63(12):1566-71. 4. Breast cancer: Incidence, mortality and survival, 2010. Office for National Statistics. 2012; 28:9. Accessed 31 August 2014. Available from: http://www.ons.gov.uk/ons/rel/cancer-unit/breast-cancer-in-england/2010/sum-1.html. 5. Worm J, Kirkin AF, Dzhandzhugazyan KN, Guldberg P. Methylation-dependent silencing of the reduced folate carrier gene in inherently methotrexate-resistant human breast cancer cells. J Biol Chem. 2001;276(43):39990-40000. 6. Thangapazham RL, Singh AK, Sharma A, Warren J, Gaddipati JP, Maheshwari RK. Green tea polyphenols and its constituent epigallocatechin gallate inhibit proliferation of human breast cancer cells in vitro and in vivo. Cancer Lett. 2007;245(1–2):232-41. 7. Stuart EC, Jarvis RM, Rosengren RJ. In vitro mechanism of action for the cytotoxicity elicited by the combination of epigallocatechin gallate and raloxifene in MDA-MB-231 cells. Oncol Rep. 2010;24(3):779-85. 8. Lecumberri E, Deupertuis YM, Mralbell R, Pichard C. Green tea polyphenol epigallocatechin-3-gallate (EGCG) as adjuvant in cancer therapy. Clin Nutr. 2013(32):894-903. 9. Dhanasekaran S. Biswal BK, Sumantran VN, Verma RS. Augmented sensitivity to methotrexate by curcumin induced overexpression of folate receptor in KG-1 cells. Biochimie 2013;95(8): 1567-73. 10. Denizot F, Lang R. Rapid colorimetric assay for cell growth and survival. modifications to the tetrazolium dye procedure giving improved sensitivity and reliability. J Immunol Methods. 1986;89(2):271-7. 11. Magee PJ, Owusu-Apenten R, McCann MJ, Gill CI, Rowland IR. (2012). Chickpea (Cicer arietinum) and other plant derived protease inhibitor concentrates inhibit breast and prostate cancer cell Proliferation in vitro. Nutri Cancer 2012; 64(5): 741-748. 12. Chien SY, Wu YC, Chung JG, Yang JS, Lu HF, Tsou MF, et al. Quercetin-induced apoptosis acts through mitochondrial- and caspase-3-dependent pathways in human breast cancer MDA-MB-231 cells. Hum Exp Toxicol. 2009;28(8):493-503. 13. Murakami A, Ashida H, Terao J. Multi-targeted cancer prevention by quercetin. Cancer Lett. 2008;269(2):315-25. 14. Wu Z, Shah A, Patel N, Yuan X. Development of methotrexate proline prodrug to overcome resistance by MDA-MB-231 cells. Bioorg Med Chem Lett. 2010;20(17):5108-12. 15. Calabrese EJ, Cancer biology and hormesis: Human tumor cell lines commonly display hormetic (biphasic) dose responses. Crit Rev Toxic. 2005;35(6):463-582. 16. Babich H, Schuck AG, Weisburg JH, Zuckerbraun HL. Research strategies in the study of the pro-oxidant nature of polyphenol nutraceuticals. J Toxicol. 2011;2011:467305.

PY - 2015/3/31

Y1 - 2015/3/31

N2 - Aim: To determine the effect of tea polyphenols and methotrexate on viability and reactive oxygen species (ROS) in a naturally resistant breast cancer cell line MDA-MB-231.Methodology: MDA-MB-231 cells were selected as a model for methotrexate resistant breast cancer. Drug tests were performed over 72 hours at concentrations 0-100 µM. Pre-treatments were with quercetin (QE) or epigallocatechin gallate (EGCG) for 5 hours followed by methotrexate. Cytotoxicity was measured using the MTT assay or resazurin fluorescence assay. ROS was determined using the 2’, 7’-dichlorofluorescein diacetate assay. Intracellular GSH was measured using the monochlorobimane assay.Results: Methotrexate was cytotoxic to MDA-MB-231 cells with IC50 of 35±4 µM. The IC50 value was 68±9.4 µM with QE and 83±16 µM for EGCG. The pre-treatment with QE and EGCG lowered the IC50 for methotrexate by 28% (P =0.009) and 16% (P=0.2027). Intracellular ROS concentrations increased after treatment with methotrexate, QE or EGCG singly and ROS decreased with combination treatment compared with the response for methotrexate only. There were no significant changes in intracellular GSH. Conclusion: Pretreatment with tea polyphenols partially sensitized breast cancer cells towards methotrexate and decreases intracellular ROS. More research is needed to optimize the sensitizing effect of tea phenols on the breast cancer cell response to methotrexate.

AB - Aim: To determine the effect of tea polyphenols and methotrexate on viability and reactive oxygen species (ROS) in a naturally resistant breast cancer cell line MDA-MB-231.Methodology: MDA-MB-231 cells were selected as a model for methotrexate resistant breast cancer. Drug tests were performed over 72 hours at concentrations 0-100 µM. Pre-treatments were with quercetin (QE) or epigallocatechin gallate (EGCG) for 5 hours followed by methotrexate. Cytotoxicity was measured using the MTT assay or resazurin fluorescence assay. ROS was determined using the 2’, 7’-dichlorofluorescein diacetate assay. Intracellular GSH was measured using the monochlorobimane assay.Results: Methotrexate was cytotoxic to MDA-MB-231 cells with IC50 of 35±4 µM. The IC50 value was 68±9.4 µM with QE and 83±16 µM for EGCG. The pre-treatment with QE and EGCG lowered the IC50 for methotrexate by 28% (P =0.009) and 16% (P=0.2027). Intracellular ROS concentrations increased after treatment with methotrexate, QE or EGCG singly and ROS decreased with combination treatment compared with the response for methotrexate only. There were no significant changes in intracellular GSH. Conclusion: Pretreatment with tea polyphenols partially sensitized breast cancer cells towards methotrexate and decreases intracellular ROS. More research is needed to optimize the sensitizing effect of tea phenols on the breast cancer cell response to methotrexate.

KW - Breast cancer

KW - methotrexate

KW - quercetin

KW - epigallocatechin gallate

KW - oxidative stress

KW - anticancer effect

M3 - Article

VL - 2

SP - 152

EP - 159

JO - Journal of Applied Life Sciences International

T2 - Journal of Applied Life Sciences International

JF - Journal of Applied Life Sciences International

SN - 2394-1103

IS - 4

ER -