Regulation of Caspase Activity During Apoptosis Induced by Baicalein in HL60 Human Leukemia Cell Line

  • Byun, Boo-Hyeong (College of Oriental Medicine, Daegu Haany University) ;
  • Kim, Bu-Yeo (Department of Medical Research, Korea Institute of Oriental Medicine)
  • Published : 2008.12.31

Abstract

Baicalein, one of the major flavonoid in Scutellaria baicalensis, has been known for its effects on proliferation and apoptosis of many tumor cell lines. Most biological effects of baicalein are thought to be from its antioxidant and prooxidant activities. In this report, baicalein was found to induce apoptosis in HL60 human promyelocytic leukemia cell line. Baicalein treatment induced DNA fragmentation and typical morphological features of apoptosis. To elucidate the mechanism of baicalein-induced apoptosis, the activities of the members of caspase family were measured. Interestingly caspase 2, 3, and 6 were significantly activated whereas caspase 1, 8, and 9 were not activated, suggesting selective involvement of specific caspases. Further, treatment with caspase inhibitors also supports the involvement of caspase 2 in apoptosis process. Although it has been reported that baicalein can induce apoptosis through many caspase pathways, the present study indicates that caspase 2 not caspase 9 pathway may be the important step in apoptosis on HL60 cell line.

References

  1. Vermeulen K, Van Bockstaele DR, Berneman ZN. Apoptosis: Mechanisms and relevance in cancer. Ann. Hematol. 84: 627-639 (2005) https://doi.org/10.1007/s00277-005-1065-x
  2. So FV, Guthrie N, Chambers AF, Carroll KK. Inhibition of proliferation of estrogen receptor-positive MCF-7 human breast cancer cells by flavonoids in the presence and absence of excess estrogen. Cancer Lett. 112: 127-133 (1997) https://doi.org/10.1016/S0304-3835(96)04557-0
  3. Yano H, Mizoguchi A, Fukuda K, Haramaki M, Ogasawara S, Momosaki S, Kojiro M. The herbal medicine Sho-saiko-to inhibits proliferation of cancer cell lines by inducing apoptosis and arrest at the G0/G1 phase. Cancer Res. 54: 448-454 (1994)
  4. Wang J, Yu Y, Hashimoto F, Sakata Y, Fujii M, Hou DX. Baicalein induces apoptosis through ROS-mediated mitochondrial dysfunction pathway in HL-60 cells. Int. J. Mol. Med. 14: 627-632 (2004)
  5. Zhao Y, Li H, Gao Z, Gong Y, Xu H. Effects of flavonoids extracted from Scutellaria baicalensis Georgi on hemin-nitrite-$H_2O_2$ induced liver injury. Eur. J. Pharmacol. 536: 192-199 (2006) https://doi.org/10.1016/j.ejphar.2006.02.045
  6. Deschamps JD, Kenyon VA, Holman TR. Baicalein is a potent in vitro inhibitor against both reticulocyte 15-human and platelet 12-human lipoxygenases. Bioorg. Med. Chem. 14: 4295-4301(2006) https://doi.org/10.1016/j.bmc.2006.01.057
  7. Woo KJ, Lim JH, Suh SI, Kwon YK, Shin SW, Kim SC, Choi YH, Park JW, Kwon TK. Differential inhibitory effects of baicalein and baicalin on LPS-induced cyclooxygenase-2 expression through inhibition of $C/EBP{\beta}$ DNA-binding activity. Immunobiology 211: 359-368 (2006) https://doi.org/10.1016/j.imbio.2006.02.002
  8. Kim BY, Han MJ, Chung AS. Effects of reactive oxygen species on proliferation of Chinese hamster lung fibroblast (V79) cells. Free Radical Bio. Med. 30: 686-698 (2001) https://doi.org/10.1016/S0891-5849(00)00514-1
  9. Datta R, Banach D, Kojima H, Talanian RV, Alnemri ES, Wong WW, Kufe DW. Activation of the CPP32 protease in apoptosis induced by 1-${\beta}$-D-arabinofuranosylcytosine and other DNA-damaging agents. Blood 88: 1936-1943 (1996)
  10. Kimuya Y, Kubo M, Tani T, Arichi S, Okuda H. Studies on Scutellariae radix. IV. Effects on lipid peroxidation in rat liver. Chem. Pharm. Bull. 29: 2610-2617 (1981) https://doi.org/10.1248/cpb.29.2610
  11. Tang W, Eisenbrand G. Chinese Drugs of Plant Origin: Chemistry, Pharmacology, and Use in Traditional and Modern Medicine. Springer Verlag, Berlin, Germany. p. 919 (1992)
  12. Li QB, You Y, Chen ZC, Lu J, Shao J, Zou P. Role of baicalein in the regulation of proliferation and apoptosis in human myeloma RPMI8226 cells. Chin. Med. J. 119: 948-952 (2006)
  13. Yoon YS, Kim JW, Kang KW, Kim YS, Choi KH, Joe CO. Poly(ADP-ribosyl)ation of histone H1 protein correlates with internucleosomal DNA fragmentation during apoptosis. J. Biol. Chem. 271: 9129-9134 (1996) https://doi.org/10.1074/jbc.271.15.9129
  14. Li YC, Tyan YS, Kuo HM, Chang WC, Hsia TC, Chung JG. Baicalein induced in vitro apoptosis undergo caspases activity in human promyelocytic leukemia HL-60 cells. Food Chem. Toxicol. 42: 37-43 (2004) https://doi.org/10.1016/j.fct.2003.08.014
  15. Huang WH, Lee AR, Chien PY, Chou TC. Synthesis of baicalein derivatives as potential anti-aggregatory and anti-inflammatory agents. J. Pharm. Pharmacol. 57: 219-225 (2005) https://doi.org/10.1211/0022357055371
  16. Liu XH, Kirschenbaum A, Yao S, Lee R, Holland JF, Levine AC. Inhibition of cyclooxygenase-2 suppresses angiogenesis and the growth of prostate cancer in vivo. J. Urol. 164: 820-825 (2000) https://doi.org/10.1016/S0022-5347(05)67321-1
  17. Makino T, Tsubouchi R, Murakami K, Haneda M, Yoshino M. Generation of reactive oxygen species and induction of apoptosis of HL60 cells by ingredients of traditional herbal medicine, Sho-saiko-to. Basic Clin. Pharmacol. Toxicol. 98: 401-405 (2006) https://doi.org/10.1111/j.1742-7843.2006.pto_328.x
  18. Prasad V, Chandele A, Jagtap JC, Sudheer Kumar P, Shastry P. ROS-triggered caspase 2 activation and feedback amplification loop in ${\beta}$-carotene-induced apoptosis. Free Radical Bio. Med. 41: 431-442 (2006) https://doi.org/10.1016/j.freeradbiomed.2006.03.009
  19. Motoo Y, Sawabu N. Antitumor effects of saikosaponins, baicalin, and baicalein on human hepatoma cell lines. Cancer Lett. 86: 91-95 (1994) https://doi.org/10.1016/0304-3835(94)90184-8
  20. Ahn HC, Lee SY, Kim JW, Son WS, Shin CG, Lee BJ. Binding aspects of baicalein to HIV-1 integrase. Mol. Cell 12: 127-130 (2001)
  21. Hwang JM, Tseng TH, Tsai YY, Lee HJ, Chou FP, Wang CJ, Chu CY. Protective effects of baicalein on tert-butyl hydroperoxide-induced hepatic toxicity in rat hepatocytes. J. Biomed. Sci. 12: 389-397 (2005) https://doi.org/10.1007/s11373-005-1572-8
  22. Sekiya K, Okuda H. Selective inhibition of platelet lipoxugenase by baicalein. Biochem. Bioph. Res. Co. 105: 1090-1095 (1982) https://doi.org/10.1016/0006-291X(82)91081-6
  23. Vistica DT, Skehan P, Scudiero DA, Monks A, Pittman A, Boyd MR. Tetrazolium-based assays for cellular viability: A critical examination of selected parameters affecting formazan production. Cancer Res. 51: 2515-2520 (1991)
  24. Ma Z, Otsuyama K, Liu S, Abroun S, Ishikawa H, Tsuyama N, Obata M, Li FJ, Zheng X, Maki Y, Miyamoto K, Kawano MM. Baicalein, a component of Scutellaria radix from Huang-Lian-Jie-Du-Tang (HLJDT), leads to suppression of proliferation and induction of apoptosis in human myeloma cells. Blood 105: 3312-3318 (2005) https://doi.org/10.1182/blood-2004-10-3915