Tumor Biology

, Volume 35, Issue 6, pp 5409–5415 | Cite as

Oxymatrine triggers apoptosis by regulating Bcl-2 family proteins and activating caspase-3/caspase-9 pathway in human leukemia HL-60 cells

  • Jun Liu
  • Yazhou Yao
  • Huifang Ding
  • Renan ChenEmail author
Research Article


With the objective of identifying promising antitumor agents for human leukemia, we carried out to determine the anticancer ability of oxymatrine on the human leukemia HL-60 cell line. In vitro experiments demonstrated that oxymatrine reduced the proliferation of HL-60 cells in a dose- and time-dependent manner via the induction of apoptosis and cell cycle arrest at G2/M and S phases. The proteins involved in oxymatrine-induced apoptosis in HL-60 cells were also examined using Western blot. The increase in apoptosis upon treatment with oxymatrine was correlated with downregulation of anti-apoptotic Bcl-2 expression and upregulation of pro-apoptotic Bax expression. Furthermore, oxymatrine induced the activation of caspase-3 and caspase-9 and the cleavage of poly(ADP-ribose) polymerase (PARP) in HL-60 cells. In addition, pretreatment with a specific caspase-3 (Z-DEVD-FMK) or caspase-9 (Z-LEHD-FMK) inhibitor significantly neutralized the pro-apoptotic activity of oxymatrine in HL-60 cells, demonstrating the important role of caspase-3 and caspase-9 in this process. Taken together, these results indicated that oxymatrine-induced apoptosis may occur through the activation of the caspase-9/caspase-3-mediated intrinsic pathway. Therefore, oxymatrine may be a potential candidate for the treatment of human leukemia.


Oxymatrine Apoptosis HL-60 Caspase Bax/Bcl-2 


Conflicts of interest



  1. 1.
    Wu SS, Chen LG, Lin RJ, Lin SY, Lo YE, Liang YC. Cytotoxicity of (−)-vitisin B in human leukemia cells. Drug Chem Toxicol. 2013;36:313–9.CrossRefPubMedGoogle Scholar
  2. 2.
    Löwenberg B, Downing JR, Burnett A. Acute myeloid leukemia. N Engl J Med. 1999;341:1051–62.CrossRefPubMedGoogle Scholar
  3. 3.
    Kang SH, Jeong SJ, Kim SH, Kim JH, Jung JH, Koh W, et al. Icariside II induces apoptosis in U937 acute myeloid leukemia cells: role of inactivation of STAT3-related signaling. PloS one. 2012;7:e28706.PubMedCentralCrossRefPubMedGoogle Scholar
  4. 4.
    Nau KC, Lewis WD. Multiple myeloma: diagnosis and treatment. Am Fam Physician. 2008;78:853–9.PubMedGoogle Scholar
  5. 5.
    Fotoohi AK, Assaraf YG, Moshfegh A, Hashemi J, Jansen G, Peters GJ, et al. Gene expression profiling of leukemia T-cells resistant to methotrexate and 7-hydroxymethotrexate reveals alterations that preserve intracellular levels of folate and nucleotide biosynthesis. Biochem Pharmacol. 2009;77:1410–7.CrossRefPubMedGoogle Scholar
  6. 6.
    Jemal A, Murray T, Samuels A, Ghafoor A, Ward E, Thun MJ. Cancer statistics, 2003. CA Cancer J Clin. 2003;53:5–26.CrossRefPubMedGoogle Scholar
  7. 7.
    Zaini RG, Brandt K, Clench MR, Le Maitre CL. Effects of bioactive compounds from carrots (Daucus carota L.), polyacetylenes, beta-carotene and lutein on human lymphoid leukaemia cells. Anticancer Agents Med Chem. 2012;12:640–52.CrossRefPubMedGoogle Scholar
  8. 8.
    Niu YP, Li LD, Wu LM. Beta-aescin: a potent natural inhibitor of proliferation and inducer of apoptosis in human chronic myeloid leukemia K562 cells in vitro. Leuk Lymphoma. 2008;49:1384–91.CrossRefPubMedGoogle Scholar
  9. 9.
    Wargovich MJ, Woods C, Hollis DM, Zander ME. Herbals, cancer prevention and health. J Nutr. 2001;131:3034S–6.PubMedGoogle Scholar
  10. 10.
    Boon H, Wong J. Botanical medicine and cancer: a review of the safety and efficacy. Expert Opin Pharmacother. 2004;5:2485–501.CrossRefPubMedGoogle Scholar
  11. 11.
    Dong Y, Xi H, Yu Y, Wang Q, Jiang K, Li L. Effects of oxymatrine on the serum levels of T helper cell 1 and 2 cytokines and the expression of the S gene in hepatitis B virus S gene transgenic mice: a study on the anti-hepatitis B virus mechanism of oxymatrine. J Gastroenterol Hepatol. 2002;17:1299–306.CrossRefPubMedGoogle Scholar
  12. 12.
    Liu J, Liu Y, Klaassen CD. The effect of Chinese hepato-protective medicines on experimental liver injury in mice. J Ethnopharmacol. 1994;42:183–91.CrossRefPubMedGoogle Scholar
  13. 13.
    Cao YG, Jing S, Li L, Gao JQ, Shen ZY, Liu Y, et al. Antiarrhythmic effects and ionic mechanisms of oxymatrine from Sophora flavescens. Phytother Res. 2010;24:1844–9.CrossRefPubMedGoogle Scholar
  14. 14.
    Cui X, Wang Y, Kokudo N, Fang D, Tang W. Traditional Chinese medicine and related active compounds against hepatitis B virus infection. Biosci Trends. 2010;4:39–47.PubMedGoogle Scholar
  15. 15.
    Deng ZY, Li J, Jin Y, Chen XL, Lü XW. Effect of oxymatrine on the p38 mitogen-activated protein kinases signalling pathway in rats with CCl4 induced hepatic fibrosis. Chin Med J (Engl). 2009;122:1449–54.Google Scholar
  16. 16.
    Zeng Z, Wang GJ, Si CW. Basic and clinical study of oxymatrine on HBV infection. J Gastroenterol Hepatol. 1999;14:295–7.Google Scholar
  17. 17.
    Chen XS, Wang GJ, Cai X, Yu HY, Hu YP. Inhibition of hepatitis B virus by oxymatrine in-vivo. World J Gastroenterol. 2001;7:49–52.PubMedGoogle Scholar
  18. 18.
    Xiang X, Wang G, Cai X, Li Y. Effect of oxymatrine on murine fulminant hepatitis and hepatocyte apoptosis. Chin Med J. 2002;115:593–6.PubMedGoogle Scholar
  19. 19.
    Lu LG, Zeng MD, Mao YM, Li JQ, Wan MB, Li CZ, et al. Oxymatrine therapy for chronic hepatitis B: a randomized double-blind and placebo-controlled multi-center trial. World J Gastroenterol. 2003;9:2480–3.PubMedGoogle Scholar
  20. 20.
    Lu LG, Zeng MD, Mao YM, Fang JY, Song YL, Shen ZH, et al. Inhibitory effect of oxymatrine on serum hepatitis B virus DNA in HBV transgenic mice. World J Gastroenterol. 2004;10:1176–9.PubMedGoogle Scholar
  21. 21.
    Mao YM, Zeng MD, Lu LG, Wan MB, Li CZ, Chen CW, et al. Capsule oxymatrine in treatment of hepatic fibrosis due to chronic viral hepatitis: a randomized, double blind, placebo-controlled, multicenter clinical study. World J Gastroenterol. 2004;10:3269–73.PubMedGoogle Scholar
  22. 22.
    Fan H, Li L, Zhang X, Liu Y, Yang C, Yang Y, et al. Oxymatrine downregulates TLR4, TLR2, MyD88, and NF-kappaB and protects rat brains against focal ischemia. Mediat Inflamm. 2009;2009:704706. doi: 10.1155/2009/704706.CrossRefGoogle Scholar
  23. 23.
    Liu XY, Fang H, Yang ZG, Wang XY, Ruan LM, Fang DR, et al. Matrine inhibits invasiveness and metastasis of human malignant melanoma cell line A375 in vitro. Int J Dermatol. 2008;47:448–56.CrossRefPubMedGoogle Scholar
  24. 24.
    Luo C, Zhu Y, Jiang T, Lu X, Zhang W, Jing Q, et al. Matrine induced gastric cancer MKN45 cells apoptosis via increasing pro-apoptotic molecules of Bcl-2 family. Toxicology. 2007;229:245–52.CrossRefPubMedGoogle Scholar
  25. 25.
    Ma L, Wen S, Zhan Y, He Y, Liu X, Jiang J. Anticancer effects of the Chinese medicine matrine on murine hepatocellular carcinoma cells. Planta Med. 2008;74:245–51.CrossRefPubMedGoogle Scholar
  26. 26.
    Zhang Y, Zhang H, Yu P, Liu Q, Liu K, Duan HY, et al. Effects of matrine against the growth of human lung cancer and hepatoma cells as well as lung cancer cell migration. Cytotechnology. 2009;59:191–200.PubMedCentralCrossRefPubMedGoogle Scholar
  27. 27.
    Bao JL, Lu JJ, Chen XP, Wang Y. Research progress in the anti-tumor effects and mechanisms of matrine and oxymatrine. Tradis Chin Drug Res Clin Pharmacol. 2012;3:369–73.Google Scholar
  28. 28.
    Song G, Luo Q, Qin J, Wang L, Shi Y, Sun C. Effects of oxymatrine on proliferation and apoptosis in human hepatoma cells. Colloids Surf B: Biointerfaces. 2006;48:1–5.CrossRefPubMedGoogle Scholar
  29. 29.
    Zhang MJ, Huang J. Recent research progress of anti-tumor mechnism matrine. Zhongguo Zhong Yao Za Zhi. 2004;29:115–8.PubMedGoogle Scholar
  30. 30.
    Jin Y, Hu J, Wang Q, Li Z, Chen Y. Effects of oxymatrine on the apoptosis of human esophageal carcinoma Eca109 cell line and its mechanism. J Huazhong Univ Sci Technol Med Sci. 2008;28:314–6.CrossRefPubMedGoogle Scholar
  31. 31.
    Zhang Y, Wang Q, Wang T, Zhang H, Tian Y, Luo H, et al. Inhibition of human gastric carcinoma cell growth in vitro by a polysaccharide from Aster tataricus. Int J Biol Macromol. 2012;51:509–13.CrossRefPubMedGoogle Scholar
  32. 32.
    Zheng L, He M, Long M, Blomgran R, Stendahl O. Pathogen-induced apoptotic neutrophils express heat shock proteins and elicit activation of human macrophages. J Immunol. 2004;173:6319–26.CrossRefPubMedGoogle Scholar
  33. 33.
    Tayarani-Najaran Z, Mousavi SH, Vahdati-Mashhadian N, Emami SA, Parsaee H. Scutellaria litwinowii induces apoptosis through both extrinsic and intrinsic apoptotic pathways in human promyelocytic leukemia cells. Nutr Cancer. 2012;64(1):80–8.CrossRefPubMedGoogle Scholar
  34. 34.
    Choi BH, Kim W, Wang QC, Kim DC, Tan SN, Yong JW, et al. Kinetin riboside preferentially induces apoptosis by modulating Bcl-2 family proteins and caspase-3 in cancer cells. Cancer Lett. 2008;261:37–45.CrossRefPubMedGoogle Scholar
  35. 35.
    Lik F, Kumar A, Bhushan S, Khan S, Bhatia A, Suri KA, et al. Reactive oxygen species generation and mitochondrial dysfunction in the apoptotic cell death of human myeloid leukemia HL-60 cells by a dietary compound withaferin A with concomitant protection by N-acetyl cysteine. Apoptosis. 2007;12:2115–33.CrossRefGoogle Scholar
  36. 36.
    Buendia B, Santa-Maria A, Courvalin JC. Caspase-dependent proteolysis of integral and peripheral proteins of nuclear membranes and nuclear pore complex proteins during apoptosis. J Cell Sci. 1999;112:1743–53.PubMedGoogle Scholar
  37. 37.
    Kerr JF, Wyllie AH, Currie AR. Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer. 1972;26:239–57.PubMedCentralCrossRefPubMedGoogle Scholar
  38. 38.
    Hengartner MO. The biochemistry of apoptosis. Nature. 2000;407:770–6.CrossRefPubMedGoogle Scholar
  39. 39.
    Kim JH, Choi YW, Park C, Jin CY, Lee YJ, da Park J, et al. Apoptosis induction of human leukemia U937 cells by gomisin N, a dibenzocyclooctadiene lignan, isolated from Schizandra chinensis Baill. Food Chem Toxicol. 2010;48:807–13.CrossRefPubMedGoogle Scholar
  40. 40.
    Kim YS, Jin SH, Lee YH, Kim SI, Park JD. Ginsenoside Rh2 induces apoptosis independently of Bcl-2, Bcl-xL, or Bax in C6Bu-1 cells. Arch Pharm Res. 1999;22:448–53.CrossRefPubMedGoogle Scholar
  41. 41.
    Ahmad N, Feyes DK, Nieminen AL, Agarwal R, Mukhtar H. Green tea constituent epigallocatechin-3-gallate and induction of apoptosis and cell cycle arrest in human carcinoma cells. J Natl Cancer Inst. 1997;89:1881–6.CrossRefPubMedGoogle Scholar
  42. 42.
    Bhalla K, Ibrado AM, Tourkina E, Tang C, Mahoney ME, Huang Y. Taxol induces internucleosomal DNA fragmentation associated with programmed cell death in human myeloid leukemia cells. Leukemia. 1993;7:563–8.PubMedGoogle Scholar
  43. 43.
    Sharma RA, Gescher AJ, Steward WP. Curcumin: the story so far. Eur J Cancer. 2005;41:1955–68.CrossRefPubMedGoogle Scholar
  44. 44.
    Su YT, Chang HL, Shyue SK, Hsu SL. Emodin induces apoptosis in human lung adenocarcinoma cells through a reactive oxygen species dependent mitochondrial signaling pathway. Biochem Pharmacol. 2005;70:229–41.CrossRefPubMedGoogle Scholar
  45. 45.
    Hockenbery DM, Oltvai ZN, Yin XM, Milliman CL, Korsmeyer SJ. Bcl-2 functions in an antioxidant pathway to prevent apoptosis. Cell. 1993;75:241–51.CrossRefPubMedGoogle Scholar
  46. 46.
    Fadeel B, Orrneius S. Apoptosis: a basic biological phenomenon with wide-ranging implications in human disease. J Intern Med. 2005;258:479–517.CrossRefPubMedGoogle Scholar
  47. 47.
    Kekre N, Griffin C, McNulty J, Pandey S. Pancratistatin causes early activation of caspase-3 and the flipping of phosphatidylserine followed by rapid apoptosis specifically in human lymphoma cells. Cancer Chemother Pharmacol. 2005;56:29–38.CrossRefPubMedGoogle Scholar
  48. 48.
    Salvesen GS, Dixit VM. Caspase activation: the induced-proximity model. Proc Natl Acad Sci U S A. 1999;96:10964–7.PubMedCentralCrossRefPubMedGoogle Scholar
  49. 49.
    Chang JT, Chen YL, Yang HT, Chen CY, Cheng AJ. Differential regulation of telomerase activity by six telomerase subunits. Eur J Biochem. 2002;269:3442–50.CrossRefPubMedGoogle Scholar
  50. 50.
    Allen RT, Hunter 3rd WJ, Agrawal DK. Morphological and biochemical characterization and analysis of apoptosis. J Pharmacol Toxicol Methods. 1997;37:215–28.CrossRefPubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2014

Authors and Affiliations

  1. 1.Department of Geriatrics, Tangdu Hospitalthe Fourth Military Medical UniversityXi’anChina
  2. 2.Department of Haematologic and RheumatismBaoji Central HospitalBaoJiChina
  3. 3.Department of HaematologyShengli Oilfield Central HospitalDongyingChina
  4. 4.Department of Haematology, Tangdu Hospitalthe Fourth Military Medical UniversityXi’anChina

Personalised recommendations