Journal of Acupuncture Research

Korean Acupuncture & Moxibustion Medicine Society (대한침구의학회)
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Bee Venom Inhibits PC-3 Cell Proliferation Through Induction of Apoptosis Via Inactivation of NF-${\kappa}B$

Bee Venom이 NF-${\kappa}B$의 불활성화에 의한 세포자멸사를 통해 PC-3 세포의 증식에 미치는 영향

  • Oh, Hyun-Jun (Dept. of Acupuncture & Moxibustion, College of Oriental Medicine, Kyungwon University) ;
  • Song, Ho-Sueb (Dept. of Acupuncture & Moxibustion, College of Oriental Medicine, Kyungwon University)
  • 오현준 (경원대학교 한의과대학 침구학교실) ;
  • 송호섭 (경원대학교 한의과대학 침구학교실)
  • Received : 2010.03.05
  • Accepted : 2010.05.26
  • Published : 2010.06.20
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Abstract

목적 : 이 연구는 봉약침의 봉독과 그 주요성분인 멜리틴이 NF-${\kappa}B$의 활성억제와 세포자멸사 관련 단백질의 발현 조절을 통하여 세포자멸사를 유도함으로써 전립선 암세포주인 PC-3 세포의 성장을 억제하는지를 확인하고 해당 기전을 살펴보고자 하였다. 방법 : 봉독이나 멜리틴을 처리한 후 PC-3의 성장억제를 관찰하기 위해 WST-1 assay, CCK-8 assay를 시행하였고, 세포자멸사 조절단백질의 변동 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}B$의 활성 변화를 관찰하기 위해 EMSA를 시행하였으며, PC-3에서 봉독이나 멜리틴과 NF-${\kappa}B$의 상호작용을 관찰하기 위해 transient transfection assay를 시행하여 세포생존율과 NF-${\kappa}B$의 활성 변동을 측정하였다. 결과 : PC-3 세포에 봉독이나 멜리틴을 처리한 후, 전립선암세포의 성장, 세포자멸사의 유발, 세포자멸사 관련 단백질의 발현, NF-${\kappa}B$의 활성, NF-${\kappa}B$의 p50, $IKK{\alpha}$, $IKK{\beta}$ 치환 후 NF-${\kappa}B$의 활성과 PC-3 세포 증식에 미치는 영향을 관찰하여 다음과 같은 결과를 얻었다. 1. PC-3 세포에서 봉독이나 멜리틴을 처리한 후 세포자멸사가 유도되어 세포성장이 억제되었고, 세포자멸사 관련 단백질 중 분리된 PARP, caspase-3, -9는 유의한 증가를, Bcl-2, XIAP, cXIAP2는 유의한 감소를 나타내었다. 2. PC-3 세포에서 봉독이나 멜리틴을 처리한 후 NF-${\kappa}B$의 활성은 유의한 감소를 나타내었다. 3. PC-3 세포에서 NF-${\kappa}B$의 p50, $IKK{\alpha}$, $IKK{\beta}$를 치환하여 작용기를 없애고 봉독이나 멜리틴을 처리하였을 경우에도 NF-${\kappa}B$의 활성이 유의한 감소를 나타내었다. 결론 : 이상의 결과는 봉독이나 멜리틴이 NF-${\kappa}B$의 활성 억제를 통하여 인간 전립선암세포주인 PC-3의 세포자멸사를 유발함으로써 증식억제 효과가 있음을 입증한 것으로, 전립선암의 예방과 치료에 대한 효과적인 치료제 개발에 도움이 될 것으로 기대된다.

Keywords

References

  1. H Gronberg. Prostate cancer epidemiology. Lancet. 2003 ; 361 : 859-64. https://doi.org/10.1016/S0140-6736(03)12713-4
  2. Walsh PC, Partin AW, Epstein JI. Cancer control and quality of life following anatomical radical retropubic prostatectomy. J Uro. 1994 ; 152 : 1831-6.
  3. Dawson N, Moul J, Higano C. Hormone refractory prostate cancer : Current issues and treatment options. Glenview IL : Physicians & Scientists Publishing Co Inc. 1999 ; 45.
  4. Stamey TA, McNeal JE. Campbell's urology, 6th ed. Philadelphia : WB Saunders. 1992 : 345.
  5. Kozlowski JM, Ellis WJ, Grayhack JT. Advanced prostatic carcinoma. Early versus late endocrine therapy. Urol Clin North Am. 1991 ; 18 : 15-24.
  6. Isaacs JT, Lundmo PI, Berges R, Martikainen P, Kyprianou N, English HF. Androgen regulation of programmed death of normal and malignant prostatic cells. J Androl. 1992 ; 13 : 457-64.
  7. Ismail M, Gomella LG. Current treatment of advanced prostate cancer. Tech Urol. 1997 ; 3 : 16-24.
  8. Green DR. Apoptotic pathways : Paper wraps stone blunts scissors. Cell. 2000 ; 102 : 1-4. https://doi.org/10.1016/S0092-8674(00)00003-9
  9. Raj GV, Sekula JA, Guo R, Madden JF, Daaka Y. Lysophosphatidic acid promotes survival of androgen-insensitive prostate cancer PC3 cells via activation of NF-kappaB. Prostate. 2004 ; 61 : 10513.
  10. Syrovets T, Gschwend JE, Buchele B et al. Inhibition of IkappaB kinase activity by acetylboswellic acetylboswellic acids promotes apoptosis in androgenindependent PC-3 prostate cancer cells in vitro and in vivo. J Biol Chem. 2005 ; 280 : 6170-80.
  11. Cattaneo-Pangrazzi RMC, Schott H, Wunderli- Allenspach H, Rothen-Rutishauser B, Guenthert M, Schwendener RA. Cell cycle arrest and p53-independent induction of apoptosis in vitro by the new anticancer drugs 5-FdUrd-PFdCydOct and dCyd-Pam-P-FdUrd in DU-145 human prostate cancer cells. J Cancer Res Clin Oncol. 2000 ; 126 : 247-56. https://doi.org/10.1007/s004320050339
  12. Lowe SW, Lin AW. Apoptosis in cancer. Carcinogenesis 2000 ; 21 : 485-95. https://doi.org/10.1093/carcin/21.3.485
  13. HS Lee, HS Song. Bee venom inhibits LNCaP cell proliferation through induction of apoptosis via inactivation of $NF-{\kappa}B$. Journal of acupuncture and moxibustion 2007 ; 25(2) : 59-74.
  14. DJ Son, MH Park, SJ Chae et al. Inhibitory effect of snake venom toxin from Vipera lebetina turanica on hormone-refractory human prostate cancer cell growth : induction of apoptosis through inactivation of nuclear factor ${\kappa}B$. Mol Cancer Ther. 2007 ; 6(2) : 675-83. https://doi.org/10.1158/1535-7163.MCT-06-0328
  15. Wolf BB, Green DR. Suicidal tendencies : apoptotic cell death by caspase family proteinases. J Biol Chem. 1999 ; 274 : 20049-52. https://doi.org/10.1074/jbc.274.29.20049
  16. American Cancer Society. Cancer facts and figures 2001. Am Cancer Soc. 2001 ; 42.
  17. DG Bostwick, J Qian, Effect of androgen deprivation therapy on prostatic intraepithelial neoplasia. Urology. 2001 ; 58 : 91-3. https://doi.org/10.1016/S0090-4295(01)01248-1
  18. BJ Feldman, D Feldman, The development of androgen-independent prostate cancer. Nat Rev Cancer. 2001 ; 1 : 34-45. https://doi.org/10.1038/35094009
  19. Lowe SW, Lin AW. Apoptosis in cancer. Carcinogenesis. 2000 ; 21 : 48595.
  20. Guseva NV, Taghiyev AF, Rokhlin OW, Cohen MB. Death receptor-induced cell death in prostate cancer. J Cell Biochem. 2004 ; 91 : 7099.
  21. Gurumurthy S, Vasudevan KM, Rangnekar VM. Regulation of apoptosis in prostate cancer. Cancer Metastasis Rev. 2001 ; 20 : 22543.
  22. Kantoff PW. New agents in the therapy of hormone-refractory patients with prostate cancer. Semin Oncol 1995 ; 22(1 Suppl 1) : 324.
  23. Isaacs JT. Apoptosis: Translating theory to therapy for prostate cancer. J Natl Cancer Inst. 2000 ; 92 : 1367-9. https://doi.org/10.1093/jnci/92.17.1367
  24. Ripple GH, Wilding G. Drug development in prostate cancer. Semin Oncol. 1999 ; 26 : 217-26.
  25. Green DR. Apoptotic pathways : Paper wraps stone blunts scissors. Cell. 2000 ; 102 : 1-4. https://doi.org/10.1016/S0092-8674(00)00003-9
  26. Thornberry NA and Lazbrik Y. Caspases : enemies within. Science. 1998 ; 281 : 1312-6. https://doi.org/10.1126/science.281.5381.1312
  27. Darmon AJ, Nicholson DW and Bleackley RC. Activation of the apoptotic protease CPP32 by cytotoxic T cell derived granzyme B Nature. 1995 ; 377 : 446-8. https://doi.org/10.1038/377446a0
  28. Sedlak TW, Oltvai ZN, Yang E et al. Multiple Bcl2 family members demonstrate selective dimerization with Bax. Proc Natl Acad Sci USA. 1995 ; 92 : 7834-8. https://doi.org/10.1073/pnas.92.17.7834
  29. Green DR and Reed JC. Mitochondria and Apoptosis. Science. 1998 ; 281 : 1309-12. https://doi.org/10.1126/science.281.5381.1309
  30. LaCasse EC, Baird S, Korneluk RG, MacKenzie AE. The inhibitors of apoptosis (IAPs) and their emerging role in cancer. Oncogene. 1998 ; 17 : 3247-59.
  31. Deveraux QL, Reed JC. IAP family proteinssuppressors of apoptosis. Genes Dev. 1999 ; 13 : 239-52. https://doi.org/10.1101/gad.13.3.239
  32. Li Y, Sarkar FH. Inhibition of nuclear factor kappaB activation in PC3 cells by genistein is mediated via Akt signaling pathway. Clin Cancer Res. 2002 ; 8 : 2369-77.