The Journal of Internal Korean Medicine (대한한방내과학회지)

The Society of Internal Korean Medicine (대한한방내과학회)
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Comparative Study of Korean Mistletoe Lectin and Bee Venom on the Anti-Cancer Effect and Its Mechanisms of Action in Hepatocellular Carcinoma Cells

상기생과 봉독이 간암 세포주 Hep G2에 대해 미치는 항암 기전 비교

  • Kim, Sung-Uk (3rd Dept. of Internal Medicine, College of Oriental Medicine, Dong-Guk University) ;
  • Kim, Bo-Ram (3rd Dept. of Internal Medicine, College of Oriental Medicine, Dong-Guk University) ;
  • Heo, Kyung (3rd Dept. of Internal Medicine, College of Oriental Medicine, Dong-Guk University) ;
  • Lim, Seong-Woo (3rd Dept. of Internal Medicine, College of Oriental Medicine, Dong-Guk University)
  • 김승욱 (동국대학교 한의과대학 비계내과학교실) ;
  • 김보람 (동국대학교 한의과대학 비계내과학교실) ;
  • 허경 (동국대학교 한의과대학 비계내과학교실) ;
  • 임성우 (동국대학교 한의과대학 비계내과학교실)
  • Published : 2009.12.30
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Abstract

Background and Objectives : Korean mistletoe lectin (Viscum album coloratum agglutinin, VCA) and bee venom (BV) have been reported to induce apoptosis in various cancer cell lines in vitro and to show antitumor activity against a variety of tumors in animal models. However, the comparative effect of VCA and BV on the anti-cancer effect and mechanisms of action has not been determined. In this study, the effect in a human hepatocellular carcinoma cell line, Hep G2 cells, was examined. Methods : Cytotoxic effects of VCA and BV on Hep G2 cells were determined by 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay in litro. The apoptotic cell death was then confirmed by propidium iodide staining and DNA fragmentation analysis. The mechanisms of action were examined by the expression of anti-apoptotic proteins and activation of mitogen-activated protein kinases. The involvement of kinase was examined in VCA or BV-induced apoptosis by using kinase inhibitors. Results : VCA and BV killed Hep G2 cells in a time and dose-dependent manner. Treatment of Hep G2 cells with VCA activated poly (ADP-ribose) polymerase-1 (PARP-1) known as a marker of apoptosis, and mitogen-activated protein kinases signaling pathways including MAPK/ERK, p38 MAPK and JNK. BV also activated PARP-1, MAPK/ERK. and p38 MAPK but not JNK. The expression level of anti-apoptotic molecule, Bcl-X, was decreased by VCA treatment but not by BV. Finally, the phosphorylation level of ERM proteins involved in the cytoskeleton homeostasis was decreased by both stimuli. VCA-induced apoptosis was partially inhibited by in the presence of JNK and p38 inhibitor, but BV only by p38 inhibitor. Conclusions : VCA-induced apoptosis is dependent on the activation of p38 and JNK. while BV-induced apoptosis is mediated by p38 activation in Hep G2 cells.

Keywords

References

  1. 辛民敎. 原色臨床本草學. 서울: 南山堂; 1991, p. 233-8.
  2. Stein GM, Schetzel M, Bussing A. Mistletoe in immunology and the clinic(short review), Anticancer Research 1998:18:3247-9.
  3. Stein GM, Pfuller U, Schietzel M. Viscotoxin-free aqueous exyracts from European mistletoe (Viscum album L.) stimulate activity of human granulocytes. Anticancer Research 1999:19:2925-8.
  4. 신민교, 송오준, 황금택. 상기생 및 곡기생 전탕액이 마우스의 항종양 면역반응에 미치는 영향. 본초분과학회지. 1994;9(1):25-47.
  5. Park RK, Kim MS, So HS, Jung BH, Moon SR, Chung SY, Ko CB, Kim BR, Chung HT. Activation of c-Jun N-terminal kinase 1 (JNK1) in mistletoe lectin II-induced apoptosis of human myeloleukemic U937 cells. Biochemical Pharmacology. 2000;60(11):1685-91. https://doi.org/10.1016/S0006-2952(00)00482-2
  6. Stoss M, Peter E, Gorter RW. Decrease of Activated Lymphocytes Four and Nine Hours after a Subcutaneous Injection of a Viscum album L. Extract in Healthy Volunteers. Nat Immun. 1998;16(5-6):185-97. https://doi.org/10.1159/000069445
  7. Fischer S, Scheffler A, Kabelitz D. Oligoclonal in vitro response of CD4 T cells to vesicles of mistletoe extracts in mistletoe-treated cancer patients. Cancer Immunol Immunother. 1997;44(3):150-6. https://doi.org/10.1007/s002620050367
  8. Deregnaucourt C, Schrevel J. Bee venom phospholopase A2 induces stage-specific growth arrest of the intraerythrocytic Plasmodium falciparum via modifications of human serum components. J Biol Chem. 2000;275(51):39973-80. https://doi.org/10.1074/jbc.M006712200
  9. Han HJ, Lee JH, Park SH, Choi HJ, Yang IS, Mar WC, Kang SK, Lee HJ. Effect of Bee Venom and Its Melittin on Apical Transporters of Renal Proximal Tubule Cells. Kidney & blood pressure research. 2000;23(6):393-9. https://doi.org/10.1159/000025988
  10. Lee J, Lim W, Eun SY, Kim SJ, Kim J. Inhibition of apamin-sensitive K+ current by hypoxia in adult rat adrenal chromaffin cells. Pflugers Arch. 2000;439(6):700-4. https://doi.org/10.1007/s004240050995
  11. von Garnier C, Astori M, Kettner A, Dufour N, Heusser C, Corradin G, Spertini F. Allegen-derived long peptide immunotherapy down-regulates specific IgE response and protects from anaphylaxis. Eur J Immunol. 2000;30(6):1638-45. https://doi.org/10.1002/1521-4141(200006)30:6<1638::AID-IMMU1638>3.0.CO;2-R
  12. 윤형석, 김용석, 이재동. 통증 관련 봉독연구에 대한 고찰. 대한약침학회지. 2000;3(1):157-75.
  13. 權奇祿, 高炯均. 봉약침요법의 면역반응에 관한 임상적 연구. 대한침구학회지. 2000:17(1):169-74.
  14. 朴贊烈, 南相水, 金昌煥, 李裁東, 姜成吉, 李潤浩, 安秉哲. 藥鍼用峰毒액이 黑色腫細胞에 미치는 抗癌效果에 對한 分子生物學的 硏究. 東西醫學硏究所論文集. 2001:2000:183-200.
  15. 박찬열, 서정철, 최도용, 안병철. 봉독약침의 항암효과에 대한 분자생물학적 연구. 대한약침학회지. 2000;3(1):1-19.
  16. Millward-Sadler SJ, Wright MO, Davies LW, Nuki G, Salter DM. Mechanotransduction via integrins and interleukin-4 results in altered aggrecan and matrix metalloproteinase 3 gene expression in normal, but not osteoarthritic, human articular chondrocytes. Arthritis Rheum. 2000;43(9):2091-9. https://doi.org/10.1002/1529-0131(200009)43:9<2091::AID-ANR21>3.0.CO;2-C
  17. Yasin B, Pang M, Turner JS, Cho Y, Dinh NN. Waring AJ, Lehrer RI, Wagar EA. Evaluation of the inactivation of infectious Herpes simplex Virus by host-defense Peptides. European Journal of Clinical Microbiology & Infectious Diseases. 2000;19(3):187-94. https://doi.org/10.1007/s100960050457
  18. Branas P, Jordan R, Fry-Smith A, Burls A, Hyde C. Treatments for fatigue in multiple sclerosis : a rapid and systematic review. Health Technol Assess. 2000;4(27):1-61.
  19. Gerst JE, Salomon Y. Inhibition by melittin and fluphenazine of melanotropin receptor function and adenylate cyclase in M2R melanoma cell membranes. Endocrinology. 1987;121(5):1766-72. https://doi.org/10.1210/endo-121-5-1766
  20. 공현식, 박동원, 류기원, 류봉하. 韓國韓醫學의 腫瘍硏究에 對한 現況;(1980年 以後의 雜誌에 報告된 論文을 中心으로). 대한한방종양학회지. 1995;1(1):55-81.
  21. Kovacs E, Hajto T, Hostanska K. Improvement of DNA repair in lymphocytes of breast cancer patients treated with Viscum album extract (Iscador). Eur J Cancer. 1991:27(12):1672-6. https://doi.org/10.1016/0277-5379(91)90443-H
  22. Bocci V. Mistletoe (viscum album) lectins as cytokine inducers and immunoadjuvant in tumor therapy. A review. J Biol Regul Homeost Agents. 1993;7(1):1-6.
  23. Krammer PH, Behrmann I, Daniel P, Dhein J, Debatin KM. Regulation of apoptosis in the immune system. Curr Opin Immunol. 1994;6:279-89. https://doi.org/10.1016/0952-7915(94)90102-3
  24. Igney FH, Krammer PH, Death and anti-death: tumour resistance to apoptosis, Nature Reviews Cancer 2. 2002;2:277-88. https://doi.org/10.1038/nrc776
  25. Janssen O, Scheffler A, Kabelitz D. In vitro effects of mistletoe extracts and mistletoe lectins. Cytotoxicity towards tumor cells due to the induction of programmed cell death (apoptosis). Arzneimittelforschung. 1993;43(11):1221-7.
  26. Kimurab C, Zhaoa QL, Kondo T, Amatsu M, Fujiwara Y. Mechanism of UV-Induced Apoptosis in Human Leukemia Cells: Roles of Ca2+/Mg2+-Dependent Endonuclease, Caspase-3, and Stress-Activated Protein Kinases. Experimental Cell Research. 1998;239:411-22. https://doi.org/10.1006/excr.1997.3912
  27. Xia Z, Dickens M, Raingeaud J, Davis RJ, Greenberg ME. Opposing effects of ERK and JNK-p38 MAP kinases on apoptosis. Science. 1995:270:1326-31. https://doi.org/10.1126/science.270.5240.1326
  28. Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C. A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nature Medicine. 2003:9:1180-6. https://doi.org/10.1038/nm911
  29. Chiarugi A. Poly(ADP-ribose) polymerase: killer or conspirator? The 'suicide hypothesis' revisited? Trends in pharmacological sciences. 2002;23(3):122-9. https://doi.org/10.1016/S0165-6147(00)01902-7
  30. Matsuzawa A, Ichijo H. Redox control of cell fate by MAP kinase: physiological roles of ASK1-MAP kinase pathway in stress signaling. Biochim Biophys Acta. 2008;1780(11):1325-36. https://doi.org/10.1016/j.bbagen.2007.12.011
  31. Kondo T, Takeuchi K, Doi Y, Yonemura S, Nagata S, Tsukita S. ERM (ezrin/radixin/moesin) -based molecular mechanism of microvillar breakdown at an early stage of apoptosis. J Cell Biol. 1997;139(3):749-58. https://doi.org/10.1083/jcb.139.3.749