Journal of Pharmacopuncture (대한약침학회지)

KOREAN PHARMACOPUNCTURE INSTITUTE (대한약침학회)
권호 표지
DOI QR코드

DOI QR Code

Bee Venom Inhibits Prostate Cancer Growth in LNCaP Xenografts via Apoptosis

Bee venom의 세포자멸사를 통한 전립선 암세포의 성장 및 LNCaP의 이종이식에 미치는 영향

  • Yang, Chang-Yeol (Dept. of Acupuncture & Moxibustion, College of Oriental Medicine, Kyungwon University) ;
  • Song, Ho-Sueb (Dept. of Acupuncture & Moxibustion, College of Oriental Medicine, Kyungwon University)
  • 양창열 (경원대학교 한의과대학 침구학교실) ;
  • 송호섭 (경원대학교 한의과대학 침구학교실)
  • Published : 2010.03.31
Download PDF
⟨ Previous Next ⟩

Abstract

연구목적 : 이 연구는 봉약침의 봉독이 NF-${\kappa}B$ 활성억제와 안드로겐 수용체 조절 단백질 및 세포자멸사 조절 단백질의 발현을 통하여 세포자멸사를 유도하고, 전립선 암세포를 이식한 쥐에서의 세포자멸사 유도 효과를 확인함으로써, 봉약침의 봉독이 생체 내에서도 세포자멸사를 유도하여 전립선암에 효과를 나타냄을 확인하고자 하였다. 실험방법 : 세포자멸사의 관찰에는 DAPI, TUNEL staining assay를 시행하였으며, 세포자멸사 조절 단백질의 변동 관찰에는 western blot analysis를 시행하였고, 세포자멸사와 연관된 NF-${\kappa}B$의 활성 변화를 관찰하기 위해 EMSA를 시행하였다. 결과 : 1. DAPI, TUNEL staining assay 결과 봉독 및 melittin을 처리한 LNCaP 세포 모두에서 세포자멸사 유도율이 유의한 증가를 나타내었다. 2. LNCaP 세포에 봉독이나 melittin을 처리한 결과, 안드로겐 수용체 조절 단백질 중 p-Akt, COX-2, calpain은 봉독과 melittin 모두에서 유의한 감소를 나타내었고, Akt는 melittin에서 유의한 감소를 나타냈으며, 봉독에서 증가하는 경향을 보였고, MMP-9은 증가하였다. 3. 생체 내에서의 봉독의 항암효과를 확인하기 위해 전립선암세포가 이식된 쥐에 봉독을 처리한 후 암세포의 부피와 무게, 쥐의 체중을 측정한 결과, 봉독을 처리한 군에서 암 세포 부피비율 및 무게는 감소하였고, 쥐의 체중은 증가하였다. 4. 전립선암세포가 이식된 쥐에 봉독을 처리한 결과, NF-${\kappa}B$ 활성에서 유의한 감소를 나타내었다. 5. 전립선암세포가 이식된 쥐에 봉독을 처리한 결과, 세포자멸사 조절 단백질 중 Bax/Bcl-2, p53, caspase-3, caspase-9, calpain은 유의한 증가를, COX-2는 유의한 감소를 나타냈으며, MMP-9는 증가를 나타내었다. 결론 : 이상의 결과는 봉독이 시험관 내에서 뿐만 아니라 생체 내에서도 NF-${\kappa}B$의 활성을 억제하고 안드로겐 수용체 조절 단백질 및 세포자멸사 조절 단백질의 조절을 통하여 인간 전립선암 세포주인 LNCaP의 세포자멸사를 유도함으로써 전립선암 세포 증식억제 효과 및 호르몬 비의존적인 전립선암으로의 전이를 지연시키는 경향이 있을 것으로 사료되고, 봉독이 전립선암의 예방과 치료에 효과적으로 활용될 수 있을 것으로 기대된다.

Keywords

References

  1. American Cancer Society. Cancer facts and figures 2001. Am Cancer Soc 2001; 42 p. CeramideTherapy in Prostate Cancer 391
  2. Dawson N, Moul J, Higano C. Hormonerefractory prostate cancer: Current issues and treatment options. Glenview, IL: Physicians & Scientists Publishing Co., Inc; 1999. 45 p.
  3. Isaacs JT. Apoptosis: Translating theory to therapy for prostate cancer. J Natl Cancer Inst 2000;92:1367-1369 https://doi.org/10.1093/jnci/92.17.1367
  4. Ripple GH, Wilding G. Drug development in prostate cancer. Semin Oncol 1999;26:217-226
  5. Green DR. Apoptotic pathways: Paper wraps stone blunts scissors. Cell 2000;102:1-4 https://doi.org/10.1016/S0092-8674(00)00003-9
  6. 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-P-FdCydOct and dCyd-Pam-P-FdUrd in DU-145 human prostate cancer cells. J Cancer Res Clin Oncol 2000;126:247-256 https://doi.org/10.1007/s004320050339
  7. Lowe SW, Lin AW. Apoptosis in cancer. Carcinogenesis 2000; 21:485-495 https://doi.org/10.1093/carcin/21.3.485
  8. Wang XZ, Beebe JR, Pwiti L, Bielawska A, Smyth MJ. Aberrant sphingolipid signaling is involved in the resistance of prostate cancer cell lines to chemotherapy. Cancer Res 1999;59:5842-5848
  9. Isaacs JT, Furuya Y, Berges R. The role of androgen in the regulation of programmed cell death/apoptosis in normal and malignant prostatic tissue. Semin Cancer Biol 1994;5:391-400
  10. Tang DG, Porter AT. Target to apoptosis: A hopeful weapon for prostate cancer. Prostate 1997;32:284-293 https://doi.org/10.1002/(SICI)1097-0045(19970901)32:4<284::AID-PROS9>3.0.CO;2-J
  11. 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
  12. Huggins C, Hodges CV. Studies on prostate cancer. I. The effects of castration, of estrogen and of androgen injection on serum phosphatases in metastatic carcinoma of the prostate. Cancer Res 1941;1:293-297
  13. Miyamoto H, Messing EM, Chang C. Androgen deprivation therapy for prostate cancer: Current status and future prospects. Prostate 2004;61:332-353 https://doi.org/10.1002/pros.20115
  14. Feldman, B.J., Feldman, D., 2001. The development of androgen independent prostate cancer. Nat. Rev. Cancer 1, 34-45 https://doi.org/10.1038/35094009
  15. Hiroshi Miyamoto, Saleh Altuwaijri, Yi Cai, Edward M. Messing, and Chawnshang Chang. Inhibition of the Akt, cyclooxygenase-2, and matrix metalloproteinase-9 pathways in combination with androgen deprivation therapy: potential therapeutic approaches for prostate cancer. Molecular carcinogenesis (2005);44:1-10 https://doi.org/10.1002/mc.20121
  16. Chang C, Saltzman A, Yeh S, Young W, Keller E, Lee HJ, Wang C, Mizokami A. Androgen receptor: An overview. Crit Rev Eukaryot Gene Expr (1995) 5:97-125 https://doi.org/10.1615/CritRevEukarGeneExpr.v5.i2.10
  17. Yang H, Murthy S, Sarkar FH, Sheng S, Reddy GP, Dou QP. Calpain-mediated androgen receptor breakdown in apoptotic prostate cancer cells. J Cell Physiol (2008);217(3):569-76 https://doi.org/10.1002/jcp.21565
  18. Lowe SW, Lin AW. Apoptosis in cancer. Carcinogenesis 2000;21:48595
  19. Guseva NV, Taghiyev AF, Rokhlin OW, Cohen MB. Death receptor-induced cell death in prostate cancer. J Cell Biochem 2004;91:7099
  20. Gurumurthy S, Vasudevan KM, Rangnekar VM. Regulation of apoptosis in prostate cancer. Cancer Metastasis Rev 2001;20:22543
  21. Kantoff PW. New agents in the therapy of hormone-refractory patients with prostate cancer. Semin Oncol 199522 (1 Suppl 1):324
  22. Jemal A, Siegel R, Ward E, Murray T, Xu J, Thun MJ. 2007. Cancer statistics, 2007. CA Cancer J Clin 57:43-66 https://doi.org/10.3322/canjclin.57.1.43
  23. Heinlein CA, Chang C. 2004. Androgen receptor in prostate cancer. Endocr Rev 25:276-308 https://doi.org/10.1210/er.2002-0032
  24. Chang C, Saltzman A, Yeh S, Young W, Keller E, Lee HJ, Wang C, Mizokami A. 1995. Androgen receptor: An overview. Crit Rev Eukaryot Gene Expr 5:97-125 https://doi.org/10.1615/CritRevEukarGeneExpr.v5.i2.10
  25. Tamura K, Furihata M, Tsunoda T, Ashida S, Takata R, Obara W, Yoshioka H, Daigo Y, Nasu Y, Kumon H, Konaka H, Namiki M, Tozawa K, Kohri K, Tanji N, Yokoyama M, Shimazui T, Akaza H, Mizutani Y,Miki T, Fujioka T, Shuin T, Nakamura Y, Nakagawa H. 2007. Molecular features of hormonerefractory prostate cancer cells by genomewide gene expression profiles. Cancer Res 67:5117-5125 https://doi.org/10.1158/0008-5472.CAN-06-4040
  26. W Zhu, PE Murtha, CY Young, Calpain inhibitor-induced apoptosis in human prostate adenocarcinoma cells. Biochem Biophys Res Commun (1995) 214:1130-7 https://doi.org/10.1006/bbrc.1995.2403
  27. Simon,H.U., Haj-Heyia,A. and Levi Schaffer, F. (2000) Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis, 5, 415-418 https://doi.org/10.1023/A:1009616228304
  28. Wyllie,A.H. (1995) The genetic regulation of apoptosis. Curr. Opin. Genet.Dev., 5(1), 97-104 https://doi.org/10.1016/S0959-437X(95)90060-8
  29. Green,D.R. and Reed,J.C. (1998) Mitochondria and Apoptosis. Science, 281,1309-1312 https://doi.org/10.1126/science.281.5381.1309
  30. Kerr JFR, Winterford CM, Harmon BV: Apoptosis. Its significance in cancer and cancer therapy. Cancer 1994;27:2013-2026
  31. Hockenbery D, Nunez G, Milliman C, Schreiber RD, Korsmeyer SJ: Bcl-2 is an inner mitochondrial membrane protein that blocks programmed cell death. Nature 1990;348:334-336 https://doi.org/10.1038/348334a0
  32. Lu QL, Abel P, Foster CS, Lalani EN: Bcl-2: Role in epithelial differentiation and oncogenesis. Hum Pathol 1996;27:102-110 https://doi.org/10.1016/S0046-8177(96)90362-7
  33. Lane DP: p53, guardian of the genome. Nature 1992;358:15-16 https://doi.org/10.1038/358015a0
  34. Kernohan NM, Cox LS: Regulation of apoptosis by Bcl-2 and its related proteins: Immunochemical challenges and therapeutic implications. J Pathol 1996;179:1-3 https://doi.org/10.1002/(SICI)1096-9896(199605)179:1<1::AID-PATH509>3.0.CO;2-E
  35. Yang E, Korsmeyer SJ: Molecular thanatopsis: A discourse on the BCL2 family and cell death. Blood 1996;88:386-401
  36. Kroemer G: The proto-oncogene Bcl-2 and its role in regulating apoptosis. Nat Med 1997;3:614-620 https://doi.org/10.1038/nm0697-614
  37. Bubendorf L, Sauter G, Moch H, Jordan P, Blochlinger A, Gasser TC, Mihatsch MJ: Prognostic significance of Bcl-2 in clinically localized prostate cancer. Am J Pathol 1996;148:1557-1565
  38. Colombel M, Symmans F, Gil S, O'Toole KM, Chopin D, Benson M, Olsson CA, Korsmeyer S, Buttyan R: Detection of the apoptosis-suppressing oncoprotein bc1-2 in hormone-refractory human prostate cancers. Am J Pathol 1993;143:390-400
  39. McDonnell TJ, Troncoso P, Brisbay SM, Logothetis C, Chung LW, Hsieh JT, Tu SM, Campbell ML: Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. Cancer Res 1992;52:6940-6944
  40. Apakama I, Robinson MC, Walter NM, Charlton RG, Fuller CE, Neal DE, Hamdy FC: bcl-2 overexpression combined with p53 protein accumulation correlates with hormone-refractory prostate cancer. Br J Cancer 1996;74:1258-1262 https://doi.org/10.1038/bjc.1996.526
  41. Hollstein M, Sidransky D, Vogelstein B, Harris CC: p53 mutations in human cancers. Science 1991;253:49-53 https://doi.org/10.1126/science.1905840
  42. Bauer JJ, Sesterhenn IA, Mostofi FK, Mcleod DG, Srivastava S, Moul JE: Elevated levels of apoptosis regulator proteins p53 and bcl-2 are independent prognostic biomarkers in surgically treated clinically localized prostate cancer. J Urol 1996;156:1511-1516 https://doi.org/10.1016/S0022-5347(01)65641-6
  43. Moul JW, Bettencourt M-C, Sesterhenn IA, Mostofi FK, McLeod DG, Sristava S, Bauer JJ: Protein expression of p53, bcl-2 and KI-67 (MIB-1) as prognostic biomarkers in patients with surgically treated, clinically localized prostate cancer. Surgery 1996;120: 159-167 https://doi.org/10.1016/S0039-6060(96)80283-2
  44. Thornberry,N.A. and Lazbrik,Y. (1998) Caspases : enemies within. Science, 281,1312-1316 https://doi.org/10.1126/science.281.5381.1312
  45. Darmon,A.J., Nicholson,D.W. and Bleackley, R.C. (1995) Activation of the apoptotic proteaseCPP32 by cytotoxic T cell derived granzyme B. Nature, 377,446-448 https://doi.org/10.1038/377446a0
  46. Stattin P, Damber J-E, Karlberg L, Bergh A: Cell proliferation assessed by KI-67 immunoreactivity on formalin fixed tissues is a predictive factor for survival in prostate cancer. J Urol 1997; 157:219-222 https://doi.org/10.1016/S0022-5347(01)65330-8
  47. Wyllie,A.H. (1995) The genetic regulation of apoptosis. Curr. Opin. Genet.Dev., 5(1), 97-104 https://doi.org/10.1016/S0959-437X(95)90060-8
  48. 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
  49. Syrovets T, Gschwend JE, Buchele B, et al. Inhibition of IkappaB kinase activity by acetyl-boswellic acids promotes apoptosis in androgen-independent PC-3 prostate cancer cells in vitro and in vivo. J Biol Chem 2005;280:617080
  50. 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;8:236977
  51. Park HJ, Lee YK, Song HS, Kim KJ, Son DJ, Lee JW and Hong JT. Melittin inhibits human prostate cancer cell growth through induction of apoptotic cell death.J. Toxicol. Pub. Health 2006;22(1):31-37.17
  52. Park HJ, Lee SH, Son DJ, Oh KW, Kim KH, Song HS, Kim GJ, Oh GT, Yoon DY, and Hong JT. Antiarthritic effect of bee venom: inhibition of Inflammation mediator generation by suppression of NF-$\kappa$B through interaction with p50 subunit. Arthritis Rheum 2004;50:3504-15 https://doi.org/10.1002/art.20626

Cited by

  1. A case report of monitoring PSA level changes in two prostate cancer patients treated with Mountain Ginseng Pharmacopuncture and Sweet Bee Venom along with western anticancer therapy vol.14, pp.4, 2011, https://doi.org/10.3831/KPI.2011.14.4.081