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도라지 유래 사포닌 platycodin D에 의한 인체 백혈병세포의 apoptosis 유도

Pro-apoptotic Effects of Platycodin D Isolated from Platycodon grandiflorum in Human Leukemia Cells

  • 박상은 (동의대학교 한의과대학 내과학교실) ;
  • 이수영 (동의대학교 한의과대학 내과학교실) ;
  • 신동역 (동의대학교 한의과대학 생화학교실) ;
  • 정진우 (동의대학교 한의과대학 생화학교실) ;
  • 진명호 (동의대학교 한의과대학 내과학교실) ;
  • 박선영 (동의대학교 한의과대학 내과학교실) ;
  • 정윤호 ((주)덕산 B&F) ;
  • 황혜진 (생활과학대학 식품영양학과) ;
  • 홍상훈 (동의대학교 한의과대학 내과학교실) ;
  • 최영현 (동의대학교 한의과대학 생화학교실)
  • Park, Sang Eun (Department of Internal Medicine, Dongeui University College of Oriental Medicine) ;
  • Lee, Su Young (Department of Internal Medicine, Dongeui University College of Oriental Medicine) ;
  • Shin, Dong Yeok (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Jeong, Jin-Woo (Department of Biochemistry, Dongeui University College of Oriental Medicine) ;
  • Jin, Myung Ho (Department of Internal Medicine, Dongeui University College of Oriental Medicine) ;
  • Park, Seon Young (Department of Internal Medicine, Dongeui University College of Oriental Medicine) ;
  • Chung, Yoon Ho (Duksan B&F Co. LTD.) ;
  • Hwang, Hye Jin (Department of Food and Nutrition, College of Human Ecology, Dongeui University) ;
  • Hong, Sang Hoon (Department of Internal Medicine, Dongeui University College of Oriental Medicine) ;
  • Choi, Yung Hyun (Department of Biochemistry, Dongeui University College of Oriental Medicine)
  • 투고 : 2013.01.23
  • 심사 : 2013.03.07
  • 발행 : 2013.03.30

초록

Platycodin D는 도라지(Platycodon grandiflorum)의 뿌리인 길경(Platycodi Radix)에 함유된 은 triterpene saponin의 일종으로 다양한 약리효능이 있는 것으로 알려져 있다. 본 연구에서는 다양한 암세포 중 platycodin D의 항암활성 감수성이 가장 높았던 인체 혈구암 U937 세포를 대상으로 platycodin D 처리에 따른 apoptosis 유발기전에 대하여 조사하였다. 본 연구의 결과에 의하면 U937 세포에서 platycodin D 처리에 의한 증식억제는 apoptosis 유발과 밀접한 연관이 있음을 알 수 있었다. 이러한 platycodin D에 의한 apoptosis 유발은 pro-apoptotic Bax의 발현 증가와 연관된 미토콘드리아 막 전위의 소실 및 caspase-3의 활성화와 기질단백질들의 분해에 의한 것임을 알 수 있었으며, IAP family 인자들의 발현 감소가 caspase의 활성 증가에도 영향을 미친 것으로 추정된다. 또한 z-DEVD-fmk 선처리에 의하여 platycodin D에 의하여 유발된 apoptosis가 유의적으로 억제되었기에, platycodin D 처리에 의하여 유발되는 apoptosis는 미토콘드리아 경로에 의하여 조절되며, caspase-3의 활성화가 핵심적인 역할을 한 것으로 생각된다.

Platycodin D is a major constituent of triterpene saponins, which is found in the root of Platycodon grandiflorum, Platycodi Radix, which is widely used in traditional Oriental medicine for the treatment of many chronic inflammatory diseases. Several pharmacological effects of this compound have been reported recently, such as anti-inflammation, immunogenicity, anti-adipogenesis, lowered cholesterol, and anti-cancer activity. However, the mechanism by which this action occurs is poorly understood. In this study, we found that platycodin D greatly increased the potential of the anti-proliferative effect in various cancer cell lines. Our data revealed that platycodin D treatment resulted in a time- and concentration-response growth inhibition of U937 cells by inducing apoptosis, as evidenced by the formation of apoptotic bodies, chromatin condensation, and the accumulation of cells in the sub-G1 phase. Apoptosis induction of U937 cells by platycodin D correlated with an increase in the Bax/Bcl-2 ratio and caused the down-regulation of IAP family members. In addition, platycodin D treatment resulted in proteolytic activation of caspase-3, the concomitant degradation of poly(ADP-ribose) polymerases, and the collapse of the mitochondria membrane potential (${\Delta}{\Psi}_m$). However, the cytotoxic effects induced by platycodin D treatment were significantly inhibited by z-DEVD-fmk, a caspase-3 inhibitor, which demonstrated the important role that caspase-3 played in the observed cytotoxic effect. These findings suggest that platycodin D may be a potential chemotherapeutic agent for use in the control of human leukemia U937 cells. These findings also provided important new insights into possible molecular mechanisms of the anti-cancer activity of platycodin D.

키워드

참고문헌

  1. Abramson, N. and Melton, B. 2000. Leukocytosis: basics of clinical assessment. Am Fam Physician 62, 2053-2060.
  2. Ahn, K. S., Noh, E. J., Zhao, H. L., Jung, S. H., Kang, S. S. and Kim, Y. S. 2005. Inhibition of inducible nitric oxide synthase and cyclooxygenase II by Platycodon grandiflorum saponins via suppression of nuclear factor-kB activation in RAW 264.7 cells. Life Sci 76, 2315-2328. https://doi.org/10.1016/j.lfs.2004.10.042
  3. Antonsson, B. and Martinou, J. C. 2000. The Bcl-2 protein family. Exp Cell Res 256, 50-57. https://doi.org/10.1006/excr.2000.4839
  4. Chang, H., Lin, H., Yi, L., Zhu, J., Zhou, Y., Mi, M. and Zhang, Q. 2010. 3,6-Dihydroxyflavone induces apoptosis in leukemia HL-60 cell via reactive oxygen species-mediated p38 MAPK/JNK pathway. Eur J Pharmacol 648, 31-38. https://doi.org/10.1016/j.ejphar.2010.08.020
  5. Choi, C. Y., Kim, J. Y., Kim, Y. S., Chung, Y. C., Seo, J. K. and Jeong, H. G. 2001. Aqueous extract isolated from platycodon grandiflorum elicits the release of nitric oxide and tumor necrosis factor-${\alpha}$ from murine macrophages. Int Immunopharmacol 1, 1141-1151. https://doi.org/10.1016/S1567-5769(01)00047-9
  6. Donovan, M. and Cotter, T. G. 2004. Control of mitochondrial integrity by Bcl-2 family members and caspase-independent cell death. Biochim Biophys Acta 1644, 133-47 https://doi.org/10.1016/j.bbamcr.2003.08.011
  7. Ghobrial, I. M., Witzig, T. E. and Adjei, A. A. 2005. Targeting apoptosis pathways in cancer therapy. CA Cancer J Clin 55, 178-194. https://doi.org/10.3322/canjclin.55.3.178
  8. Gilliland, D. G., Jordan, C. T. and Felix, C. A. 2004. The molecular basis of leukemia. Hematology Am Soc Hematol Educ Program 80-97.
  9. Han, S. I., Kim, Y. S. and Kim, T. H. 2008. Role of apoptotic and necrotic cell death under physiologic conditions. BMB Rep 41, 1-10. https://doi.org/10.5483/BMBRep.2008.41.1.001
  10. Holcik, M., Gibson, H. and Korneluk, R. G. 2001. XIAP:apoptotic brake and promising therapeutic targe. Apoptosis 6, 253-261. https://doi.org/10.1023/A:1011379307472
  11. Hussein, M. R., Haemel, A. K. and Wood, G. S. 2003. Apoptosis and melanoma: molecular mechanisms. J Pathol 199, 275-288. https://doi.org/10.1002/path.1300
  12. Jin, Z. and El-Deiry, W. S. 2005. Overview of cell death signaling pathways. Cancer Biol Ther 4, 139-163. https://doi.org/10.4161/cbt.4.2.1508
  13. Kaufmann, S. H., Desnoyers, S., Ottaviano, Y., Davidson, N. E. and Poirier, G. G. 1993. Specific proteolytic cleavage of poly(ADP-ribose) polymerase: an early marker of chemotherapy- induced apoptosis. Cancer Res 53, 3976-3985.
  14. Khan, N., Adhami, V. M. and Mukhtar, H. 2008. Apoptosis by dietary agents for prevention and treatment of cancer. Biochem Pharmacol 76, 1333-1339. https://doi.org/10.1016/j.bcp.2008.07.015
  15. Konishi, T., Tada, A., Shoji, J. and Tanaka, O. 1976. The structures of platycodin A and C, monoacetylated saponins of the roots of Platycodon grandiflorum. Chem Pharm Bull(ToKyo) 26, 668-670.
  16. Lazebnik, Y. A., Kaufmann, S. H., Desnoyers, S., Poirier, G. G. and Earnshaw, W. C. 1994. Cleavage of poly(ADP-ribose) polymerase by a proteinase with properties like ICE. Nature 371, 346-347. https://doi.org/10.1038/371346a0
  17. Lee, H., Kang, R., Kim, Y. S., Chung, S. I. and Yoon, Y. 2010. Platycodin D inhibits adipogenesis of 3T3 L1 cells by modulating kruppel like factor 2 and peroxisome proliferator activated receptor gamma. Phytother Res 24, 161-167. https://doi.org/10.1002/ptr.3054
  18. Meiler, J. and Schuler, M. 2006. Therapeutic targeting of apoptotic pathways in cancer. Curr Drug Targets 7, 1361-1369. https://doi.org/10.2174/138945006778559175
  19. Okada, H. and Mak, T. W. 2004. Pathways of apoptotic and non-apoptotic death in tumour cells. Nat Rev Cancer 4, 592-603. https://doi.org/10.1038/nrc1412
  20. Salvesen, G. S. and Duckett, C. S. 2002. IAP proteins: blocking the road to death's door. Nat Rev Mol Cell Biol 3, 401-410. https://doi.org/10.1038/nrm830
  21. Tada, A., Kanejwa, Y., Shoji, J. and Shibata, S. 1975. Studies on the saponins of the root of Platycodon grandiflorum A, De Can dolle. I. Isolation and the structure of platycodin D. Chem Pharm Bull (Tokyo) 23, 2965-2972. https://doi.org/10.1248/cpb.23.2965
  22. Wang, C., Schuller Levis, G. B., Lee, E. B., Levis, W. R., Lee, D. W., Kim, B. S., Park, S. Y. and Park, E. 2004. Platycodin D and D3 isolated from the root of Platycodon grandiflorum modulate the production of nitric oxide and secretion of TNF-${\alpha}$ in activated RAW 264.7 cells. Int Immunopharmacol 4, 1039-1049. https://doi.org/10.1016/j.intimp.2004.04.005
  23. Zhao, H. L., Cho, K. H., Ha, Y. W., Jeong, T. S., Lee, W. S. and Kim, Y. S. 2006. Cholesterol-lowering effect of platycodin D in hypercholesterolemic ICR mice. Eur J Pharmacol 537, 166-173. https://doi.org/10.1016/j.ejphar.2006.03.032

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