Journal of Acupuncture Research

Korean Acupuncture & Moxibustion Medicine Society (대한침구의학회)
권호 표지
DOI QR코드

DOI QR Code

Synergistic Effects of Bee Venom and Natural Killer Cells on B16F10 Melanoma Cell Growth Inhibition through IL-4-mediated Apoptosis

  • Sin, Dae Chul (Department of Acupuncture & Moxibustion Medicine, College of Oriental Medicine, Gachon University) ;
  • Kang, Mi Suk (Department of Acupuncture & Moxibustion Medicine, College of Oriental Medicine, Gachon University) ;
  • Song, Ho Sueb (Department of Acupuncture & Moxibustion Medicine, College of Oriental Medicine, Gachon University)
  • Received : 2017.01.11
  • Accepted : 2017.02.07
  • Published : 2017.02.20
Download PDF
⟨ Previous Next ⟩

Abstract

Objectives : We investigated the synergistic effects of bee venom (BV) and natural killer (NK) cells on B16F10 melanoma cell apoptosis mediated by IL-4. Methods : We performed a cell viability assay to determine whether BV can enhance the inhibitory effect of NK-92MI cells on the growth of B16F10 melanoma cells, and western blot analysis to detect changes in the expression of IL-4, $IL-4R{\alpha}$, and other apoptosis-related proteins. EMSA was performed to observe the activity of STAT6. To confirm that the inhibitory effect of BV and NK cells was mediated by IL-4, the above tests were repeated after IL-4 silencing by siRNA (50 nM). Results : B16F10 melanoma cells co-cultured with NK-92MI cells and simultaneously treated by BV ($5{\mu}g/ml$) showed a higher degree of proliferation inhibition than when treated by BV ($5{\mu}g/ml$) alone or co-cultured with NK-92MI cells alone. Expression of IL-4, $IL-4R{\alpha}$, and that of other pro-apoptotic proteins was also enhanced after co-culture with NK-92MI cells and simultaneous treatment with BV ($5{\mu}g/ml$). Furthermore, the expression of anti-apoptotic bcl-2 decreased, and the activity of STAT6, as well as the expression of STAT6 and p-STAT6 were enhanced. IL-4 silencing siRNA (50 nM) in B16F10 cells, the effects of BV treatment and NK-92MI co-culture were reversed. Conclusion : These results suggest that BV could be an effective alternative therapy for malignant melanoma by enhancing the cytotoxic and apoptotic effect of NK cells through an IL-4-mediated pathway.

Keywords

References

  1. McCourt C, Dolan O, Gormley G. Malignant melanoma: a pictorial review. Ulster Med J. 2014;83(2):103-10.
  2. Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin. 2010;60(5):277-300. https://doi.org/10.3322/caac.20073
  3. Dave S. B. Hoon, Marinelle B, Edward O, Donald L. Morton, Reiko F. Irie. Modulation of Human Melanoma Cells by Interleukin-4 and in combination with ${\gamma}$-Interferon or ${\alpha}$-Tumor Necrosis Factor. Cancer Res. 1991;51(8):2002-8.
  4. Obiri NI, Siegel JP, Varricchio F, Puri RK. Expression of high-affinity IL-4 receptors on human melanoma, ovarian and breast carcinoma cells. Clin Exp Immunol. 1994;95(1):148-55. https://doi.org/10.1111/j.1365-2249.1994.tb06029.x
  5. Robert L. Tepper, Paul K. Pattengale, Philp Leder. Murine Interleukin-4 display potent anti-tumor activity in vivo. Cell. 1989;57(3):503-12. https://doi.org/10.1016/0092-8674(89)90925-2
  6. Pattabhiraman S, Santosh N. IL-4 Induces Apoptosis in A549 Lung Adenocarcinoma Cells: Evidence for the Pivotal Role of 15-Hydroxyeicosatetraenoic Acid Binding to Activated Peroxisome Proliferator-Activated Receptor ${\gamma}$ Transcription Factor. J Immunol 2003;170(2):887-94. https://doi.org/10.4049/jimmunol.170.2.887
  7. Hilary S Warren, Mark J Smyth. NK cells and apoptosis. Immunology and Cell. Biology. 1999;77:64-75.
  8. S Bogdanov. Bee Venom: Composition, Health, Medicine: A Review. Swiss : Bee product Science. 2016:1-11.
  9. Park JH, Jeong YJ, Park KK et al. Melittin suppresses PMA-induced tumor cell invasion by inhibiting NF-kappaB and AP-1-dependent MMP-9 expression. Mol. Cells. 2010;29:209-15. https://doi.org/10.1007/s10059-010-0028-9
  10. Park MH, Choi MS, Kwak DH et al. Anticancer effect of bee venom in prostate cancer cells through activation of caspase pathway via inactivation of $NF-{\kappa}B$. Prostate. 2010;61:801-12.
  11. Zhenjian C, Kam-Meng TW, William N.Rom. NF-kappaB in Lung Tumorigenesis. Cancers. 2011;3(4):4258-68. https://doi.org/10.3390/cancers3044258
  12. Veronique B, Michael K. Is $NF-{\kappa}B$ a good target for cancer therapy? Hopes and pitfalls. Nature Reviews Drug Discovery. 2009;8(1):33-40. https://doi.org/10.1038/nrd2781
  13. Kollipara PS, Kim JH, Won D et al. Co-culture with NK-92MI cells enhanced the anti-cancer effect of Bee Venom on NSCLC cells by inactivation of $NF-{\kappa}B$. Archives Of Pharmacal Research. 2014;37(3):379-89. https://doi.org/10.1007/s12272-013-0319-8
  14. Choi KE, Hwang CJ, Gu SM et al. Cancer Cell Growth Inhibitory Effect of Bee Venom via Increase of Death Receptor 3 Expression and Inactivation of NF-kappa B in NSCLC Cells. Toxins. 2014;6:2210-28. https://doi.org/10.3390/toxins6082210
  15. Liu S, Yu M, He Y et al. Melittin prevents liver cancer cell metastasis through inhibition of the Rac1-dependent pathway. Hepatology. 2008;47:1964-73. https://doi.org/10.1002/hep.22240
  16. Kim JH, Song HS. Bee Venom Enhanced Cytotoxic Effect of Natural Killer Cells on Human Lung Cancer Through Inducing Extrinsic Apoptosis. The Acupunct. 2014;31(1):111-9. https://doi.org/10.13045/acupunct.2014011
  17. Sung HJ, Song HS. Synergistic Effect of Natural Killer Cells and Bee Venom on Inhibition of NCI-H157 Cell Growth. The Acupunct. 2016;33(1):47-56. https://doi.org/10.13045/acupunct.2016005
  18. Ghobrial IM, Witzig TE, Adjei AA. Targeting apoptosis pathways in cancer therapy. CACancer J Clin. 2005;55:178-94. https://doi.org/10.3322/canjclin.55.3.178
  19. Ashkenazi A. Targeting death and decoy receptors of the tumor necrosis factor superfamily. Nat Rev, Cancer. 2002;2:420-30. https://doi.org/10.1038/nrc821
  20. Timmer T, de Vries EG, de Jong S. Fas receptor-mediated apoptosis. A clinical application. J Pathol. 2002;196:125-34. https://doi.org/10.1002/path.1028
  21. Kischkel FC, Hellbardt S, Behrmann I et al. Cytotoxicity-dependent APO-1 (Fas/CD95)- associated proteins form a death-inducing signaling complex (DISC) with the receptor. EMBO J. 1995;14:5579-88.
  22. Peter ME, Krammer PH. The CD95(APO-1/Fas) DISC and beyond. Cell Death Differ. 2003;10:26-35. https://doi.org/10.1038/sj.cdd.4401186
  23. The Korean society of Pathologists. Textbook of Pathology. Seoul: KMS. 2010:22-7.
  24. Takeda K, Tanaka T, Shi W et al. Essential role of Stat6 in IL-4 signalling. Nature. 1996;380:627-30. https://doi.org/10.1038/380627a0