Abstract
Glycyrrhetinic acid (GA) is the active compound in Glycyrrhizae radix, a famous traditional Chinese medicine. Recently the anticancer activity of GA became the focus of scientific interest and many GA derivatives were developed as anti-tumor lead compounds. We previously reported that AEGA, a GA derivative, has proliferation inhibition and apoptosis-inducing activity in various human tumor cells. The present study was undertaken to further investigate the molecular mechanisms involved in AEGA-induced apoptosis in human leukemia K562 cells. AEGA can inhibit the growth of K562 cells in dose- and time-dependent manners determined by the MTT assay. Induction of apoptosis was evidenced by morphological changes and biochemical markers such as cell shrinkage, chromatin condensation and DNA ladder formation. Further mechanistic analysis revealed that AEGA induced apoptosis through the collapse of mitochondrial membrane potential, the accumulation of the cytosolic cytochrome c and the activation of caspase-9 and caspase-3. The apoptosis induction by AEGA was associated with the alteration in the ratio of Bcl-2/Bax protein expression. These results suggest that AEGA may induce apoptosis through a mitochondria-mediated pathway, and might have the therapeutic value against hematological malignancies.
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Abbreviations
- GA:
-
Glycyrrhetinic acid
- AEGA:
-
3-Acetyl-2-ene-glycyrrhetinic acid
- DMSO:
-
Dimethyl sulphoxide
- FBS:
-
Fetal bovine serum
- MTT:
-
3-(4,5-dimethylthiazoyl)-2,5-diphenyltetrazolium bromide
References
Basanez G, Zhang J, Chau BN, Maksaev GI, Frolov VA, Brandt TA, Burch J, Hardwick JM, Zimmerberg J (2001) Pro-apoptotic cleavage products of Bcl-xL form cytochrome c-conducting pores in pure lipid membranes. J Biol Chem 276:31083–31091
Chadalapaka G, Jutooru I, McAlees A, Stefanac T, Safe S (2008) Structure-dependent inhibition of bladder and pancreatic cancer cell growth by 2-substituted glycyrrhetinic and ursolic acid derivatives. Bioorg Med Chem Lett 18:2633–2639
Chintharlapalli S, Papineni S, Jutooru I, McAlees A, Safe S (2007) Structure-dependent activity of glycyrrhetinic acid derivatives as peroxisome proliferator-activated receptor gamma agonists in colon cancer cells. Mol Cancer Ther 6:1588–1598
Chintharlapalli S, Papineni S, Abdelrahim M, Abudayyeh A, Jutooru I, Chadalapaka G, Wu F, Mertens-Talcott S, Vanderlaag K, Cho SD, Smith RR, Safe S (2009) Oncogenic microRNA-27a is a target for anticancer agent methyl 2-cyano-3, 11-dioxo-18beta-olean-1, 12-dien-30-oate in colon cancer cells. Int J Cancer 125:1965–1974
Csuk R, Schwarz S, Kluge R, Strohl D (2010) Synthesis and biological activity of some antitumor active derivatives from glycyrrhetinic acid. Eur J Med Chem 45:5718–5723
Davis EA, Morris DJ (1991) Medicinal uses of licorice through the millennia: the good and plenty of it. Mol Cell Endocrinol 78:1–6
Duan H, Luan J, Liu Q, Yagasaki K, Zhang G (2010) Suppression of human lung cancer cell growth and migration by berbamine. Cytotechnology 62:341–348
Fiore C, Eisenhut M, Ragazzi E, Zanchin G, Armanini D (2005) A history of the therapeutic use of liquorice in Europe. J Ethnopharmacol 99:317–324
Gao Y, Guo X, Li X, Liu D, Song D, Xu Y, Sun M, Jing Y, Zhao L (2010) The synthesis of glycyrrhetinic acid derivatives containing a nitrogen heterocycle and their antiproliferative effects in human leukemia cells. Molecules 15:4439–4449
Gao Z, Hu J, Kang X, Xu C, Ju Y (2011) Synthesis of A-ring functional derivatives of 18β-glycyrrhetinic acid and antiproliferative effect in tumor cells. Chem J Chin Univ, 29 Aug 2011 (Epub ahead of print)
Grebenova D, Kuzelova K, Fuchs O, Halada P, Havlicek V, Marinov I, Hrkal Z (2004) Interferon-alpha suppresses proliferation of chronic myelogenous leukemia cells K562 by extending cell cycle S-phase without inducing apoptosis. Blood Cells Mol Dis 32:262–269
Gu BW, Mason PJ (2010) A new potential anti-leukemia drug from a tropical plant. Leuk Res 34:1420–1421
Heerdt BG, Houston MA, Anthony GM, Augenlicht LH (1998) Mitochondrial membrane potential (delta psi(mt)) in the coordination of p53-independent proliferation and apoptosis pathways in human colonic carcinoma cells. Cancer Res 58:2869–2875
Hu J, Wu Y, Zhao C, Ju Y (2010) Synthesis and anti-tumor activity of opened A-ring modified 18β-glycyrrhetinic acid derivatives. Chem J Chin Univ 31:1762–1768
Huang Z (2000) Bcl-2 family proteins as targets for anticancer drug design. Oncogene 19:6627–6631
Huang Y, Liu Q, Liu K, Yagasaki K, Zhang G (2009) Suppression of growth of highly-metastatic human breast cancer cells by norcantharidin and its mechanisms of action. Cytotechnology 59:209
Jutooru I, Chadalapaka G, Chintharlapalli S, Papineni S, Safe S (2009) Induction of apoptosis and nonsteroidal anti-inflammatory drug-activated gene 1 in pancreatic cancer cells by a glycyrrhetinic acid derivative. Mol Carcinog 48:692–702
Law JC, Ritke MK, Yalowich JC, Leder GH, Ferrell RE (1993) Mutational inactivation of the p53 gene in the human erythroid leukemic K562 cell line. Leuk Res 17:1045–1050
Liu P, Han ZC (2003) Treatment of acute promyelocytic leukemia and other hematologic malignancies with arsenic trioxide: review of clinical and basic studies. Int J Hematol 78:32–39
Liu D, Song D, Guo G, Wang R, Lv J, Jing Y, Zhao L (2007) The synthesis of 18beta-glycyrrhetinic acid derivatives which have increased antiproliferative and apoptotic effects in leukemia cells. Bioorg Med Chem 15:5432–5439
Liu Q, Duan H, Luan J, Yagasaki K, Zhang G (2009) Effects of theanine on growth of human lung cancer and leukemia cells as well as migration and invasion of human lung cancer cells. Cytotechnology 59:211–217
Luan J, Duan H, Liu Q, Yagasaki K, Zhang G (2010) Inhibitory effects of norcantharidin against human lung cancer cell growth and migration. Cytotechnology 62:349–355
Marchetti P, Castedo M, Susin SA, Zamzami N, Hirsch T, Macho A, Haeffner A, Hirsch F, Geuskens M, Kroemer G (1996) Mitochondrial permeability transition is a central coordinating event of apoptosis. J Exp Med 184:1155–1160
McGahon A, Bissonnette R, Schmitt M, Cotter KM, Green DR, Cotter TG (1994) BCR-ABL maintains resistance of chronic myelogenous leukemia cells to apoptotic cell death. Blood 83:1179–1187
Olson M, Kornbluth S (2001) Mitochondria in apoptosis and human disease. Curr Mol Med 1:91–122
Shao HJ, Qing C, Wang F, Zhang YL, Luo DQ, Liu JK (2005) A new cytotoxic lanostane triterpenoid from the basidiomycete Hebeloma versipelle. J Antibiot (Tokyo) 58:828–831
Shibata S (2000) A drug over the millennia: pharmacognosy, chemistry, and pharmacology of licorice. Yakugaku Zasshi 120:849–862
Tallman MS (1996) Differentiating therapy with all-trans retinoic acid in acute myeloid leukemia. Leukemia 10:S12–S15
Vahedi F, Fathi NM, Bozari K (2008) Evaluation of inhibitory effect and apoptosis induction of Zyzyphus Jujube on tumor cell lines, an in vitro preliminary study. Cytotechnology 56:105–111
Yu P, Liu Q, Liu K, Yagasaki K, Wu E, Zhang G (2009) Matrine suppresses breast cancer cell proliferation and invasion via VEGF-Akt-NF-kappaB signaling. Cytotechnology 59:219–229
Zhang G, Miura Y, Yagasaki K (1999) Effects of green, oolong and black teas and related components on the proliferation and invasion of hepatoma cells in culture. Cytotechnology 31:37–44
Zhang G, Miura Y, Yagasaki K (2001) Inhibitory effects of theanine and sera from theanine-fed rats on receptor-mediated cancer cell invasion beneath mesothelial-cell monolayers. Cytotechnology 36:195–200
Zhang Y, Zhang H, Yu P, Liu Q, Liu K, Duan H, Luan G, Yagasaki K, Zhang G (2009) Effects of matrine against the growth of human lung cancer and hepatoma cells as well as lung cancer cell migration. Cytotechnology 59:191–200
Acknowledgments
This work is supported in part by Zhejiang Provincial Natural Science Foundation of China (Y4110320), Natural Science Foundation of China (Nos. 20772071, 20972086) and the Opening Foundation of Biomedicine Engineering Key Discipline, Zhejiang, China (SWYX0901).
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Gao, Z., Kang, X., Hu, J. et al. Induction of apoptosis with mitochondrial membrane depolarization by a glycyrrhetinic acid derivative in human leukemia K562 cells. Cytotechnology 64, 421–428 (2012). https://doi.org/10.1007/s10616-011-9419-9
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DOI: https://doi.org/10.1007/s10616-011-9419-9