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Medicarpin and millepurpan, two flavonoids isolated from Medicago sativa, induce apoptosis and overcome multidrug resistance in leukemia P388 cells.
Phytomedicine. 2015 Dec 01; 22(13):1186-94.P

Abstract

BACKGROUND

High consumption of flavonoids has been associated with a decrease risk of cancer. Alfalfa (Medicago sativa) leaves have been widely used in traditional medicine and is currently used as a dietary supplement because of their high nutrient content. We previously reported the cytotoxic activity of alfalfa leaf extracts against several sensitive and multidrug resistant tumor cell lines.

HYPOTHESIS/PURPOSE

We aimed to determine whether medicarpin and millepurpan, two isoflavonoids isolated from alfalfa leaves, may have pro-apoptotic effects against drug-sensitive (P388) and multidrug resistant P388 leukemia cells (P388/DOX).

STUDY DESIGN/METHODS

Cells were incubated with medicarpin or millepurpan for the appropriate time. Cell viability was assessed by the MTT assay. DNA fragmentation was analyzed by agarose gel electrophoresis. Cell cycle analysis was realized by flow cytometry technics. Caspases 3 and 9 activities were measured using Promega caspACE assay kits. Proteins and genes expression were visualized respectively by western-blot using specific antibodies and RT-PCR assay.

RESULTS

P-glycoprotein-expressing P388/DOX cells did not show resistance to medicarpin (IC50 ≈ 90 µM for P388 and P388/DOX cells) and millepurpan (IC50 = 54 µM and 69 µM for P388 and P388/DOX cells, respectively). Treatment with medicarpin or millepurpan triggered apoptosis in sensitive as well as multidrug resistant P388 cells. These effects were mediated through the mitochondrial pathway by modifying the balance pro/anti-apoptotic proteins. While 3 µM doxorubicin alone could not induce cell death in P388/DOX cells, concomitant treatment with doxorubicin and subtoxic concentration of medicarpin or millepurpan restored the pro-apoptotic cascade. Each compound increased sensitivity of P388/DOX cells to doxorubicin whereas they had no effect in sensitive P388 cells. Vinblastine cytotoxicity was also enhanced in P388/DOX cells (IC50 = 210 nM to 23 and 25 nM with medicarpin and millepurpan, respectively). This improved sensitivity was mediated by an increased uptake of doxorubicin in P388/DOX cells expressing P-gp. P-gp expression was not altered by exposure to medicarpin and millepurpan.

CONCLUSION

These data indicate that medicarpin and millepurpan possess pro-apoptotic properties and potentiate the cytotoxicity of chemotherapy drugs in multidrug resistant P388 leukemia cells by modulating P-gp-mediated efflux of drugs. These flavonoids may be used as chemopreventive agents or as sensitizer to enhance cytotoxicity of chemotherapy drugs in multidrug resistant cancer cells.

Authors+Show Affiliations

Laboratoire de Biochimie et Biologie Moléculaire, Faculté de Pharmacie, URCA, Reims, France.Laboratoire de Pharmacognosie, Faculté de Pharmacie, SFR Cap Santé, ICMR-CNRS UMR 7312, Reims, France.Service d'endocrinologie, de diabétologie et de nutrition, CHU Robert-Debré, Reims, France.MEDyC UMR CNRS/URCA no. 7369, Faculté des Sciences, SFR Cap Santé, URCA, Reims, France.MEDyC UMR CNRS/URCA no. 7369, Faculté de Pharmacie, SFR Cap Santé, URCA, Reims, France.Laboratoire de Pharmacognosie, Faculté de Pharmacie, SFR Cap Santé, ICMR-CNRS UMR 7312, Reims, France.Laboratoire de Biochimie et Biologie Moléculaire, Faculté de Pharmacie, URCA, Reims, France. Electronic address: claudie.madoulet@univ-reims.fr.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

26598918

Citation

Gatouillat, Grégory, et al. "Medicarpin and Millepurpan, Two Flavonoids Isolated From Medicago Sativa, Induce Apoptosis and Overcome Multidrug Resistance in Leukemia P388 Cells." Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, vol. 22, no. 13, 2015, pp. 1186-94.
Gatouillat G, Magid AA, Bertin E, et al. Medicarpin and millepurpan, two flavonoids isolated from Medicago sativa, induce apoptosis and overcome multidrug resistance in leukemia P388 cells. Phytomedicine. 2015;22(13):1186-94.
Gatouillat, G., Magid, A. A., Bertin, E., El btaouri, H., Morjani, H., Lavaud, C., & Madoulet, C. (2015). Medicarpin and millepurpan, two flavonoids isolated from Medicago sativa, induce apoptosis and overcome multidrug resistance in leukemia P388 cells. Phytomedicine : International Journal of Phytotherapy and Phytopharmacology, 22(13), 1186-94. https://doi.org/10.1016/j.phymed.2015.09.005
Gatouillat G, et al. Medicarpin and Millepurpan, Two Flavonoids Isolated From Medicago Sativa, Induce Apoptosis and Overcome Multidrug Resistance in Leukemia P388 Cells. Phytomedicine. 2015 Dec 1;22(13):1186-94. PubMed PMID: 26598918.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Medicarpin and millepurpan, two flavonoids isolated from Medicago sativa, induce apoptosis and overcome multidrug resistance in leukemia P388 cells. AU - Gatouillat,Grégory, AU - Magid,Abdulmagid Alabdul, AU - Bertin,Eric, AU - El btaouri,Hassan, AU - Morjani,Hamid, AU - Lavaud,Catherine, AU - Madoulet,Claudie, Y1 - 2015/10/24/ PY - 2015/06/29/received PY - 2015/09/15/revised PY - 2015/09/24/accepted PY - 2015/11/25/entrez PY - 2015/11/26/pubmed PY - 2016/1/12/medline KW - Alfalfa KW - Apoptosis KW - Flavonoids KW - Medicarpin KW - Millepurpan KW - Multidrug resistance SP - 1186 EP - 94 JF - Phytomedicine : international journal of phytotherapy and phytopharmacology JO - Phytomedicine VL - 22 IS - 13 N2 - BACKGROUND: High consumption of flavonoids has been associated with a decrease risk of cancer. Alfalfa (Medicago sativa) leaves have been widely used in traditional medicine and is currently used as a dietary supplement because of their high nutrient content. We previously reported the cytotoxic activity of alfalfa leaf extracts against several sensitive and multidrug resistant tumor cell lines. HYPOTHESIS/PURPOSE: We aimed to determine whether medicarpin and millepurpan, two isoflavonoids isolated from alfalfa leaves, may have pro-apoptotic effects against drug-sensitive (P388) and multidrug resistant P388 leukemia cells (P388/DOX). STUDY DESIGN/METHODS: Cells were incubated with medicarpin or millepurpan for the appropriate time. Cell viability was assessed by the MTT assay. DNA fragmentation was analyzed by agarose gel electrophoresis. Cell cycle analysis was realized by flow cytometry technics. Caspases 3 and 9 activities were measured using Promega caspACE assay kits. Proteins and genes expression were visualized respectively by western-blot using specific antibodies and RT-PCR assay. RESULTS: P-glycoprotein-expressing P388/DOX cells did not show resistance to medicarpin (IC50 ≈ 90 µM for P388 and P388/DOX cells) and millepurpan (IC50 = 54 µM and 69 µM for P388 and P388/DOX cells, respectively). Treatment with medicarpin or millepurpan triggered apoptosis in sensitive as well as multidrug resistant P388 cells. These effects were mediated through the mitochondrial pathway by modifying the balance pro/anti-apoptotic proteins. While 3 µM doxorubicin alone could not induce cell death in P388/DOX cells, concomitant treatment with doxorubicin and subtoxic concentration of medicarpin or millepurpan restored the pro-apoptotic cascade. Each compound increased sensitivity of P388/DOX cells to doxorubicin whereas they had no effect in sensitive P388 cells. Vinblastine cytotoxicity was also enhanced in P388/DOX cells (IC50 = 210 nM to 23 and 25 nM with medicarpin and millepurpan, respectively). This improved sensitivity was mediated by an increased uptake of doxorubicin in P388/DOX cells expressing P-gp. P-gp expression was not altered by exposure to medicarpin and millepurpan. CONCLUSION: These data indicate that medicarpin and millepurpan possess pro-apoptotic properties and potentiate the cytotoxicity of chemotherapy drugs in multidrug resistant P388 leukemia cells by modulating P-gp-mediated efflux of drugs. These flavonoids may be used as chemopreventive agents or as sensitizer to enhance cytotoxicity of chemotherapy drugs in multidrug resistant cancer cells. SN - 1618-095X UR - https://www.unboundmedicine.com/medline/citation/26598918/Medicarpin_and_millepurpan_two_flavonoids_isolated_from_Medicago_sativa_induce_apoptosis_and_overcome_multidrug_resistance_in_leukemia_P388_cells_ L2 - https://linkinghub.elsevier.com/retrieve/pii/S0944-7113(15)00314-1 DB - PRIME DP - Unbound Medicine ER -