Journal of Korean Neurosurgical Society

The Korean Neurosurgical Society (대한신경외과학회)
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Enhancement of Arsenic Trioxide ($As_2O_3$)-Mediated Apoptosis Using Berberine in Human Neuroblastoma SH-SY5Y Cells

  • Kim, Dae-Won (Department of Neurosurgery School of Medicine Wonkwang University) ;
  • Ahan, Song-Ho (Department of Neurosurgery School of Medicine Wonkwang University) ;
  • Kim, Tae-Young (Department of Neurosurgery School of Medicine Wonkwang University)
  • Published : 2007.11.30
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Abstract

Objective : Arsenic trioxide ($As_2O_3$) has been used as an anticancer agent in traditional Chinese medicine for thousand years and berberine is an isoquinoline alkaloid present that has indicated significant antimicrobial activity. We have examined the combined anticancer effects of $As_2O_3$ and berberine against the human neuroblastoma (HNB) SH-SY5Y cells in vitro, and to elucidate underlying molecular mechanism. Methods : HNB SH-SY5Y cells were treated with $2\;{\mu}M\;As_2O_3$ and $75\;{\mu}g/ml$ berberine, and their survival, cell death mechanism as well as synergistic cytotoxic effects were estimated by using MTT assay, DAPI staining, agarose gel electrophoresis, flow cytometric analysis, and western blot analysis. Results : The combined treatment of two drugs also markedly decreased cell viability. The cytotoxic effects of two drugs were revealed as apoptosis characterized by chromatin condensation, DNA fragmentation, and the loss of mitochondrial membrane potential. The apoptotic cytotoxicity was accompanied by activation of caspase-3 protease as well as decreased the expression of Bcl-2, Bid, and Bcl-x/L. In addition, the cells treated with combination of two drugs also showed significantly increased intracellular reactive oxygen species levels and lipid peroxidation compared to cells $As_2O_3$or berberine only. Conclusion : Combined treatment of $As_2O_3$ with berberine induced activation of apoptotic signaling pathways in HNB SH-SY5Y cells. These results suggest that the possibility of the combined treatment of two chemotherapeutic agents with low concentration improving cytotoxic effect for cancer cells with minimal side effects.

Keywords

References

  1. Akihter MH, Sabir M, Bhide NK : Anti-inflammatory effect of berberine in rats injected locally with cholera toxin. Indian J Med Res 65: 133-141, 1977
  2. Amin AH, Subbaiah TV, Abbasi KM : Berberine sulfate: antimicrobial activity, bioassay, and mode of action. Can J Microbiol 15 : 1067-1076, 1969 https://doi.org/10.1139/m69-190
  3. Baumgartner M, Sturlan S, Roth E, Wessner B, Bachleitner-Hofmann T : Enhancement of arsenic trioxide-mediated apoptosis using Docosahexaenoic acid in arsenic trioxide-resistant sold tumor cells. Int J Cancer 112: 707-712, 2004 https://doi.org/10.1002/ijc.20462
  4. Evans AM, Tallman MS, Gartenhaus RB : The potential of arsenic trioxide in the treatment of malignant disease: past, present, and future. Leuk Res 28 : 891-900, 2004 https://doi.org/10.1016/j.leukres.2004.01.011
  5. Hail NJ, Lotan R : Apoptosis induction by the natural product cancer chemopreventive agent deguelin is mediated through the inhibition of mitochondrial bioenergetics. Apoptosis 9 : 437-447, 2004 https://doi.org/10.1023/B:APPT.0000031449.57551.e1
  6. Hougardy BM, van der Zee AG, van den Heuvel FA, Timmer T, de Vries EG, de Jong S, et al : Sensitivity to Fas-mediated apoptosis in high-risk HPV-positive human cervical cancer cells : Relationship with Fas, caspase-8, and Bid. Gynecol Oncol 97: 353-364, 2005 https://doi.org/10.1016/j.ygyno.2005.01.036
  7. Hwang JM, Kuo HC, Tseng TH, Liu JY, Chu CY: Berberine induces apoptosis through a mitochondria/caspases pathway in human hepatoma cells. Arch Toxicol 28 : 1-12, 2005
  8. Iizuka N, Hazama N, Yoshimura S, Yoshino K, Tangoku S, Miyamoto A, et al : Anticachectic effects of the natural herb Coptidis rhizoma and Berberine on mice bearing colon 26/clone 20 adenocarcinoma. Int J Cancer 99: 286-291, 2002 https://doi.org/10.1002/ijc.10338
  9. Jakubowicz-Gil J, Paduch R, Piersiak T, Glowniak K, Gawron A, Kanderfer-Szerszen M : The effect of quercetin on pro-apoptotic activity of cisplatin in HeLa cells. Biochem Pharmaco 69 : 1343-1350, 2005 https://doi.org/10.1016/j.bcp.2005.01.022
  10. Jantova S, Cipak L, Cernakova M, Kost'alova D: Effect of berberine on proliferation, cell cycle and apoptosis in HeLa and LI210 cells. J Pharm Pharmacol 55: 1143-1149, 2003 https://doi.org/10.1211/002235703322277186
  11. Jantova S, Cipak L, Letasiova S : Berberine induces apoptosis through a mitochondrial/caspase pathway in human promonocyric U937 cells. Toxicol In Vitro 21: 25-31, 2007 https://doi.org/10.1016/j.tiv.2006.07.015
  12. Kang YH, Lee E, Choi MK, Ku JL, Kim SH, Park YG, et al : Role of reactive oxygen species in the induction of apoptosis by ${\alpha}$-tocopheryl succinate. Int J Cancer 112: 385-392, 2004 https://doi.org/10.1002/ijc.20424
  13. Kerr JF, Wyllie AH, Currie AR : Apoptosis : a basic biological phenomenon with wide ranging implication in tissue kinetics. Br J Cancer 26: 239-257, 1972 https://doi.org/10.1038/bjc.1972.33
  14. Kong B, Huang S, Wang W, Ma D, Qu X, Jiang J, et al : Arsenic trioxide induces apoptosis in cisplatin-sensitive and -resistant ovarian cancer cell lines. Int J Gynecol Cancer 15 : 872-877, 2005 https://doi.org/10.1111/j.1525-1438.2005.00251.x
  15. Kumagai T, Shih LY, Hughes SV, Desmond JC, O'Kelly J, Hewison M, et al : 19-Nor-1,25$(OH)_2D_2$ (a novel, noncalcemic vitamin D analogue), combined with arsenic trioxide, has potent antitumor activity against myeloid leukemia. Cancer Res 65 : 2488-2497, 2005 https://doi.org/10.1158/0008-5472.CAN-04-2800
  16. Kuo CL, Chi CW, Liu TY : The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett 203: 127-137, 2004 https://doi.org/10.1016/j.canlet.2003.09.002
  17. Li XK, Motwani M, Tong W, Bommann W, Schwarts GK : Huanglian, a Chinese herbal extract, inhibits cell growth by suppressing the expression of cyclin B1 and inhibiting CDC2 kinase activity in hwnan cancer cells. Mol Pharmacol 58 : 1287-1293, 2000 https://doi.org/10.1124/mol.58.6.1287
  18. Liu Q, Hilsenbeck S, Gazitt Y: Arsenic trioxide-induced apoptosis in myeloma cells: p53-dependent G1 or G2/M cell cycle arrest, activation of caspase-8 or caspase-9, and synergy with APO2/TRAIL. Blood 101 : 4078-4087, 2003 https://doi.org/10.1182/blood-2002-10-3231
  19. Maeda H, Hori S, Ohizumi H, Segawa T, Kakehi Y, Ogawa O, et al: Effective treatment of advanced solid tumors by the combination of arsenic trioxide and L-buthioninesulfoximine. Cell Death Differ 11 : 737-746, 2004 https://doi.org/10.1038/sj.cdd.4401389
  20. Maeda S, Suqiura T, Saikawa Y, Kubota T, Otani Y, Kumai K, et al: Docetaxel enhances the cytotoxicity of cisplatin to gastric cancer cells by modification of intracellular platinum metabolism. Cancer Sci 95 : 679-684, 2004 https://doi.org/10.1111/j.1349-7006.2004.tb03329.x
  21. Nazarewicz RR, Zenebe WJ, Parihar A, Larson SK, Alidema E, Choi J, et al : T amoxifen induces oxidative stress and mitochondrial apoptosis via stimulating mitochondrial nitric oxide synthase. Cancer Res 67 : 1282-1290, 2007 https://doi.org/10.1158/0008-5472.CAN-06-3099
  22. Notarbartolo M, Poma P, Perri D, Dusonchet L, Cervello M, D'Alessandro N : Antitumor effects of curcumin, alone or combination with cisplatin or doxorubicin, on human hepatic cancer cells. Analysis of their possible relationship to changes in NF-kB activation levels and in IAP gene expression. Cancer Lett 224 : 53-65, 2005 https://doi.org/10.1016/j.canlet.2004.10.051
  23. Sauer H, Wartenberg M, Hescheler J : Reactive oxygen species as intracellula messengers during cell growth and differentiation. Cell Physiol Biochem 11 : 173-186, 2001 https://doi.org/10.1159/000047804
  24. Scholz C, Wieder T, Starck L, Essmann F, Schulze-Osthoff K, Dorken B, et al : Arsenic trioxide triggers a regulated form of caspase-independent necrotic cell death via the mitochondrial death pathway. Oncogene 24: 1904-1913, 2005 https://doi.org/10.1038/sj.onc.1208233
  25. Tada-Oikawa S, Hiraku Y, Kawanishi M, Kawanish S : Mechanism for generation of hydrogen peroxide and change of mitochondrial membrane potential during rotenone-induced apoptosis. Life Science 73 : 3277-3288, 2003 https://doi.org/10.1016/j.lfs.2003.06.013
  26. Takara K, Horibe S, Obara Y, Yoshikawa E, Ohnishi N, Yokoyama T : Effects of 19 herbal extracts on the sensitivity to paclitaxel or 5-fluorouracil in HeLa cells. Biol Pharm Bull 28 : 138-142, 2005 https://doi.org/10.1248/bpb.28.138
  27. Takeuchi H, Kondo Y, Fujiwara K, Kanzawa T, Aoki H, Mills GB, et al : Synergistic Augmentation of Rapamycin-Induced Autophagy in Malignant Glioma Cells by Phosphatidylinositol 3-Kinase/Protein Kinase B Inhibitors. Cancer Res 65 : 3336-3346, 2005 https://doi.org/10.1158/0008-5472.CAN-04-3640
  28. Vermes I, Haanen C, Steffens-Nakken H, Reutelingsperger C: A novel assay for apoptosis. Flow cyrometric detection of phosphatidylserine expression on early apoptotic cells using fluorescein labelled annexin V. J Immunol Methods 184: 39-51, 1995 https://doi.org/10.1016/0022-1759(95)00072-I