DOI QR코드

DOI QR Code

누에배양세포에서 인간형 재조합단백질 각질세포 성장인자 생산

Production of Recombinant Human Keratinocyte Growth Factor from Bombyx mori (Lepidopera: Bombycidae) Bm5 Cells

  • 한송이 (충남대학교 의학전문대학원 해부학교실) ;
  • 진초이 (충남대학교 의학전문대학원 해부학교실) ;
  • 권기상 (충남대학교 의학전문대학원 해부학교실) ;
  • 윤은영 (농촌진흥청 국립농업과학원) ;
  • 구태원 (농촌진흥청 국립농업과학원) ;
  • 김승환 (충남대병원 응급의학과) ;
  • 최종순 (한국기초과학지원연구원 생명과학연구부) ;
  • 유권 (한국생명공학연구원 장수과학연구센터) ;
  • 권오유 (충남대학교 의학전문대학원 해부학교실)
  • Han, Song-Yi (Department of Anatomy, School of Medicine, Chungnam National University) ;
  • Jin, Cho-Yi (Department of Anatomy, School of Medicine, Chungnam National University) ;
  • Kwon, Ki-Sang (Department of Anatomy, School of Medicine, Chungnam National University) ;
  • Yun, Eun-Young (Department of Sericulture & Entomology, National Institute of Agriculture Science and Technology, R.D.A.) ;
  • Goo, Tae-Won (Department of Sericulture & Entomology, National Institute of Agriculture Science and Technology, R.D.A.) ;
  • Kim, Seung-Whan (Department of Emergency Medicine, Chungnam National University Hospital) ;
  • Choi, Jong-Soon (Division of Life Science, Korea Basic Science Institute) ;
  • Yu, Kweon (Aging Research Center, Korea Research Institute of Bioscience and Biotechnology) ;
  • Kwon, O-Yu (Department of Anatomy, School of Medicine, Chungnam National University)
  • 투고 : 2011.03.03
  • 심사 : 2011.05.24
  • 발행 : 2011.06.30

초록

인간형 재조합단백질 각질세포 성장인자를 안정적으로 생산하는 누에 배양 세포(Bm5-hKGF cell)을 만들었다. 이 세포에서 분비되어 배지에 포함된 양은 15-20 ng/ml 정도였다. Bm5-hKGF cell에 누에의 PDI를 함께 발현시키면 세포외 분비량이 2배 증가하였다. Wound healing migration assay 결과 누에세포에서 생산된 인간형 재조합단백질 각질세포 성장인자는 세포생장을 촉진하는 활성을 가지고 있었다. 본 실험의 결과는 누에배양세포를 사용하여 저비용으로 양질의 인간형 재조합단백질을 대량생산 할 수 있는 것을 기대한다.

Using silkworm Bombyx mori Bm5 cells, we established a stable cell line expressing the human keratinocyte growth factor (hKGF), named by the Bm5-hKGF cell, in which the protein hKGF is synthesized in the cell and secreted in the cell culture supernatant (CCS) at approximately 15-20 ng/ml. When the Bm5-hKGF cell was co-expressed with B. mori protein disulfide isomerase (bPDI) cDNA, its secretion increased by about two times the original amount. Through wound healing migration assay, it was demonstrated that the secreted hKGF included in the CCS has a very powerful biological activity of keratinocyte proliferation. We expect to produce useful human recombinant proteins from silkworm cultured cells in large quantities at low prices.

키워드

참고문헌

  1. Athar, U. and T. C. Gentile. 2009. Keratinocyte growth factor. Expert Opin. Biol. Ther. 9, 779-787. https://doi.org/10.1517/14712590902976837
  2. Goo, T. W., E. Y. Yun, K. H. Choi, S. H. Kim, S. K. Nho, S. W. Kang, and O. Y. Kwon. 2004. ATFC is a novel transducer for the unfolded protein response in Bombyx mori BM5 cells. Biochem. Biophys. Res. Commun. 325, 626-631. https://doi.org/10.1016/j.bbrc.2004.10.069
  3. Goo, T. W., E. Y. Yun, S. W. Kim, K. H. Choi, S. W. Kang, K. Kwon, K. Yu, and O. Y. Kwon. 2008. Bombyx mori protein disulfide isomerase enhances the production of nuecin, an antibacterial protein. BMB Reports 41, 400-403. https://doi.org/10.5483/BMBRep.2008.41.5.400
  4. Hille, A., S. Grüger, H. Christiansen, H. A. Wolff, B. Volkmer, J. Lehmann, W. Dörr, and M. Rave-Frank. 2010. Effect of tumour-cell-derived or recombinant keratinocyte growth factor (KGF) on proliferation and radioresponse of human epithelial tumour cells (HNSCC) and normal keratinocytes in vitro. Radiat. Environ. Biophys. 49, 261-270. https://doi.org/10.1007/s00411-010-0271-7
  5. Hitchman, R. B., R. D. Possee, and L. A. King. 2009. Baculovirus expression systems for recombinant protein production in insect cells. Recent Pat. Biotechnol. 3, 46-54. https://doi.org/10.2174/187220809787172669
  6. Kelly, R. M., E. M. Goren, P. A. Taylor, S. N. Mueller, H. E. Stefanski, M. J. Osborn, H. S. Scott, E. A. Komarova, A. V. Gudkov, G. A. Holländer, and B. R. Blazar. 2010. Short-term inhibition of p53 combined with keratinocyte growth factor improves thymic epithelial cell recovery and enhances T-cell reconstitution after murine bone marrow transplantation. Blood 115, 1088-1097. https://doi.org/10.1182/blood-2009-05-223198
  7. Rodriguez, L. G., X. Wu, and J. L. Guan. 2005. Wound-healing assay. Methods Mol. Biol. 294, 23-29.
  8. Ryu, S. H., S. Y. Moon, Y. J. Yang, S. R. Moon, J. P. Hong, J. Choi, and S. W. Lee. 2009. Recombinant human epidermal growth factor accelerates the proliferation of irradiated human fibroblasts and keratinocytes in vitro and in vivo. J. Radiat. Res. 50, 545-552. https://doi.org/10.1269/jrr.09066