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. 2010 Dec;71(6):384–397. doi: 10.1016/S0011-393X(10)80004-9

Effects of griseofulvin on apoptosis through caspase-3- and caspase-9-dependent pathways in K562 leukemia cells: An in vitro study

Ning Zhong 1, Hankui Chen 2, Quanlin Zhao 3, Hongwei Wang 2,*, Xin Yu 4, Ashley M Eaves 2, Weihua Sheng 5, Jingcheng Miao 5, Fengmei Cui 5, Jinzhi Wang 5,*
PMCID: PMC3969622  PMID: 24688157

Abstract

Background: Griseofulvin, an oral nontoxic antifungal drug, has been reported to possess anticancer effect in human cancer cells, while the mechanisms are not completely understood.

Objective: The aim of this study was to investigate the cytotoxic effect of griseofulvin on K562 cells and to understand its underlying molecular pathways.

Methods: K562 cells were treated with griseofulvin at different concentrations for 24 hours, and the inhibition effect of griseofulvin on K562 cell proliferation was assessed by tetrazolium salt colorimetric assay. Apoptosis was assessed by examining nuclear morphology and quantifying phosphatidylserine externalization, and alterations in cellular morphology were analyzed by laser scanning confocal microscopy for fluorescent analysis. Flow cytometry was used in the analysis of cell cycle, mitochondrial membrane potential, and caspase pathways.

Results: Griseofulvin could inhibit the growth of K562 cells in a dose-dependent manner with a mean (SD) inhibitory concentration of 50% value of 15.38 (1.35) μg/mL compared with untreated controls. Apoptosis was induced in K562 cells (38.35% [2.73%]; P < 0.01) by griseofulvin with the observation of both an increase in phosphatidylserine level and accumulation of chromatin nucleation in griseofulvintreated cells. In addition, cell-cycle analysis using propidium iodide staining suggested a significant G2/M accumulation (increase from mean 17.64% [4.49%] to 48.29 [1.89%]; P < 0.01) as a result of griseofulvin treatment. Flow cytometry analysis found that griseofulvin treatment was associated with the depolarization of the mitochondrial membrane in K562 cells. Furthermore, increased activities of caspase-3 by 22.15-fold (P < 0.01) and caspase-9 by 16.73-fold (P < 0.01) were observed in K562 cells after griseofulvin treatment compared with the untreated control; a decrease of caspase-8 activity was also observed, but the change was not statistically significant.

Conclusions: These findings suggest that griseofulvin inhibited growth of K562 cells and induced cell apoptosis through cell-cycle arrest and mitochondrial membrane potential decrease as well as caspase-3 and -9 activation. Further testing is needed to evaluate the potential of griseofulvin as a candidate in the chemotherapy of hematologic malignancies.

Key words: griseofulvin, caspase-3, caspase-9, K562 cells, apoptosis

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Contributor Information

Hongwei Wang, Email: zuguodadi@msn.com.

Jinzhi Wang, Email: hwang1@medicine.bsd.uchicago.edu.

References

  • 1.Chan YC, Friedlander SF. New treatments for tinea capitis. Curr Opin Infect Dis. 2004;17:97–103. doi: 10.1097/00001432-200404000-00005. [DOI] [PubMed] [Google Scholar]
  • 2.De Carli L, Larizza L. Griseofulvin. Mutat Res. 1988;195:91–126. doi: 10.1016/0165-1110(88)90020-6. [DOI] [PubMed] [Google Scholar]
  • 3.Czymmek KJ, Bourett TM, Shao Y. Live-cell imaging of tubulin in the filamentous fungus Magnaporthe grisea treated with anti-microtubule and anti-microfilament agents. Protoplasma. 2005;225:23–32. doi: 10.1007/s00709-004-0081-3. [DOI] [PubMed] [Google Scholar]
  • 4.Jordan MA, Wilson L. Microtubules as a target for anticancer drugs. Nat Rev Cancer. 2004;4:253–265. doi: 10.1038/nrc1317. [DOI] [PubMed] [Google Scholar]
  • 5.Panda D, Rathinasamy K, Santra MK, Wilson L. Kinetic suppression of microtubule dynamic instability by griseofulvin: Implications for its possible use in the treatment of cancer. Proc Natl Acad Sci U S A. 2005;102:9878–9883. doi: 10.1073/pnas.0501821102. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 6.Ho YS, Duh JS, Jeng JH. Griseofulvin potentiates antitumorigenesis effects of nocodazole through induction of apoptosis and G2/M cell cycle arrest in human colorectal cancer cells. Int J Cancer. 2001;91:393–401. doi: 10.1002/1097-0215(200002)9999:9999<::aid-ijc1070>3.0.co;2-#. [DOI] [PubMed] [Google Scholar]
  • 7.Rebacz B, Larsen TO, Clausen MH. Identification of griseofulvin as an inhibitor of centrosomal clustering in a phenotype-based screen. Cancer Res. 2007;67:6342–6350. doi: 10.1158/0008-5472.CAN-07-0663. [DOI] [PubMed] [Google Scholar]
  • 8.Uen YH, Liu DZ, Weng MS. NF-kappa pathway is involved in griseofulvin-induced G2/M arrest and apoptosis in HL-60 cells. J Cell Biochem. 2007;101:1165–1175. doi: 10.1002/jcb.21240. [DOI] [PubMed] [Google Scholar]
  • 9.Carnero A. Targeting the cell cycle for cancer therapy. Br J Cancer. 2002;87:129–133. doi: 10.1038/sj.bjc.6600458. [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 10.Green DR, Reed JC. Mitochondria and apoptosis. Science. 1998;281:1309–1312. doi: 10.1126/science.281.5381.1309. [DOI] [PubMed] [Google Scholar]
  • 11.Shih CM, Wu JS, Ko WC. Mitochondria-mediated caspase-independent apoptosis induced by cadmium in normal human lung cells. J Cell Biochem. 2003;89:335–347. doi: 10.1002/jcb.10488. [DOI] [PubMed] [Google Scholar]
  • 12.Sen N, Das B, Ganguly B. Camptothecin induced mitochondrial dysfunction leading to programmed cell death in unicellular hemoflagellate Leishmania donovani. Cell Death Differ. 2004;11:924–936. doi: 10.1038/sj.cdd.4401435. [DOI] [PubMed] [Google Scholar]
  • 13.Reers M, Smiley ST, Mottola-Hartshorn C. Mitochondrial membrane potential monitored by JC-1 dye. Methods Enzymol. 1995;260:406–417. doi: 10.1016/0076-6879(95)60154-6. [DOI] [PubMed] [Google Scholar]
  • 14.Torres K, Horwitz SB. Mechanisms of Taxol-induced cell death are concentration dependent. Cancer Res. 1998;58:3620–3626. [PubMed] [Google Scholar]
  • 15.Murray AW. Recycling the cell cycle: Cyclins revisited. Cell. 2004;116:221–234. doi: 10.1016/s0092-8674(03)01080-8. [DOI] [PubMed] [Google Scholar]
  • 16.Thompson CB. Apoptosis in the pathogenesis and treatment of disease. Science. 1995;267:1456–1462. doi: 10.1126/science.7878464. [DOI] [PubMed] [Google Scholar]
  • 17.Pietenpol JA, Stewart ZA. Cell cycle checkpoint signaling: Cell cycle arrest versus apoptosis. Toxicology. 2002;181-182:475–481. doi: 10.1016/s0300-483x(02)00460-2. [DOI] [PubMed] [Google Scholar]
  • 18.Hsu YL, Kuo YC, Kuo PL. Apoptotic effects of extract from Antrodia camphorate fruiting bodies in human hepatocellular carcinoma cell lines. Cancer Lett. 2005;221:77–89. doi: 10.1016/j.canlet.2004.08.012. [DOI] [PubMed] [Google Scholar]
  • 19.Green D, Kroemer G. The central executioners of apoptosis: Caspases or mitochondria? Trends Cell Biol. 1998;8:267–271. doi: 10.1016/s0962-8924(98)01273-2. [DOI] [PubMed] [Google Scholar]
  • 20.Cho SG, Choi EJ. Apoptotic signaling pathways: Caspases and stress-activated protein kinases. J Biochem Mol Biol. 2002;35:24–27. doi: 10.5483/bmbrep.2002.35.1.024. [DOI] [PubMed] [Google Scholar]
  • 21.Li P, Nijhawan D, Budihardjo I. Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell. 1997;91:479–489. doi: 10.1016/s0092-8674(00)80434-1. [DOI] [PubMed] [Google Scholar]
  • 22.Liu X, Kim CN, Yang J. Induction of apoptotic program in cell-free extracts: Requirement for dATP and cytochrome c. Cell. 1996;86:147–157. doi: 10.1016/s0092-8674(00)80085-9. [DOI] [PubMed] [Google Scholar]
  • 23.Crompton M. The mitochondrial permeability transition pore and its role in cell death. Biochem J. 1999;341:233–249. [PMC free article] [PubMed] [Google Scholar]
  • 24.Pallis M, Grundy M, Turzanski J. Mitochondrial membrane sensitivity to depolarization in acute myeloblastic leukemia is associated with spontaneous in vitro apoptosis, wild-type TP53, and vicinal thiol/disulfide status. Blood. 2001;98:405–413. doi: 10.1182/blood.v98.2.405. [DOI] [PubMed] [Google Scholar]
  • 25.Robertson JD, Orrenius S. Molecular mechanisms of apoptosis induced by cytotoxic chemicals. Crit Rev Toxicol. 2000;30:609–627. doi: 10.1080/10408440008951122. [DOI] [PubMed] [Google Scholar]

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