International Journal of Industrial Entomology and Biomaterials

Korean Society of Sericultural Science (한국잠사학회)
권호 표지
DOI QR코드

DOI QR Code

Functional analysis of Bombyx mori Decapentaplegic gene for bone differentiation in a mammalian cell

  • Park, Seung-Won (Department of Biotechnology, Catholic University of Daegu) ;
  • Goo, Tae-Won (Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration) ;
  • Choi, Gwang-Ho (Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kang, Seok-Woo (Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Sung-Wan (Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration) ;
  • Kim, Seong-Ryul (Department of Agricultural Biology, National Academy of Agricultural Science, Rural Development Administration)
  • Received : 2013.08.06
  • Accepted : 2013.08.17
  • Published : 2013.09.30
Download PDF
⟨ Previous Next ⟩

Abstract

Bone morphogenetic proteins (BMPs) belong to the transforming growth factor (TGF-${\beta}$) superfamily and are involved in osteoblastic differentiation. The largest TGF-${\beta}$ superfamily subgroup shares genetic homology with human BMPs (hBMPs) and silkworm decapentaplegic (dpp). In addition, hBMPs are functionally interchangeable with Drosophila dpp. Bombyx mori dpp may induce bone formation in mammalian cells. To test this hypothesis, we synthesized the 1,285-base pairs cDNA of full-length B. mori dpp using total RNAs obtained from the fat body of 3-day-old of the $5^{th}$ instar larvae and cloned the cDNA into the pCEP4 mammalian expression vector. Next, B. mori dpp was expressed in C3H10T1/2 cells. The target cells transfected with the pCEP4-Bm dpp plasmid showed biological functions similar to those of osteogenic differentiation induction growth factors such as hBMPs. We determined the relative mRNA expression rates of Runt-related transcription factor 2 (RUNX2), osterix, osteocalcin, and alkaline phosphatase (ALP) to validate the osteoblast-specific differentiation effects of B. mori dpp by performing quantitative real-time RT-PCR. Interestingly, mRNA expression levels of the 3 marker genes except RUNX2, in cells expressing B. mori dpp were much higher than those in control cells and C3H10T1/2 cells transfected with pCEP4. These results suggested that B. mori dpp signaling regulates osterix expression during osteogenic differentiation via RUNX2-independent mechanisms.

Keywords

References

  1. Aono A, Hazama M, Notoya K, Taketomi S, Yamasaki H, Tsukuda R et al. (1995) Potent Ectopic Bone-Inducing Activity of Bone Morphogenetic Protein-4/7 Heterodimer. Biochem Biophys Res Commun 210, 670-677. https://doi.org/10.1006/bbrc.1995.1712
  2. Bilic R, Simic P, Jelic M, Stern-Padovan R, Dodig D, van Meerdervoort HP et al. (2006) Osteogenic protein-1 (BMP-7) accelerates healing of scaphoid non-union with proximal pole sclerosis. Int Orthop 30, 128-134. https://doi.org/10.1007/s00264-005-0045-z
  3. Chen Y, Riese MJ, Killinger MA, Hoffmann FM (1998) A genetic screen for modifiers of Drosophila decapentaplegic signaling identifies mutations in punt, Mothers against dpp and the BMP-7 homologue, 60A. Development 125, 1759-1768.
  4. Choi YH, Gu YM, Oh JW, Lee KY (2011) Osterix is regulated by Erk1/2 during osteoblast differentiation. Biochem Biophys Res Commun 415, 472-478. https://doi.org/10.1016/j.bbrc.2011.10.097
  5. Edelman ER, Nugent MA, Karnovsky MJ (1993) Perivascular and intravenous administration of basic fibroblast growth factor: vascular and solid organ deposition. Proc. Natl. Acad. Sci. USA 90, 1513-1517. https://doi.org/10.1073/pnas.90.4.1513
  6. Kunnapuu J, Shimmi O (2010) Evolutional imprints on the sequences of BMP2/4/DPP type proteins. Fly (Austin) 4, 21-23. https://doi.org/10.4161/fly.4.1.10652
  7. Matsubara T, Kida K, Yamaguchi A, Hata K, Ichida F, Meguro H et al. (2008) BMP2 regulates Osterix through Msx2 and Runx2 during osteoblast differentiation. J Biol Chem 283, 29119-29125. https://doi.org/10.1074/jbc.M801774200
  8. Matsumoto Y, Otsuka F, Takano M, Mukai T, Yamanaka R, Takeda M et al. (2010) Estrogen and glucocorticoid regulate osteoblast differentiation through the interaction of bone morphogenetic protein-2 and tumor necrosis factor-alpha in C2C12 cells. Mol Cell Endocrinol 325, 118-127. https://doi.org/10.1016/j.mce.2010.05.004
  9. Nakashima K, Zhou X, Kunkel G, Zhang Z, Deng JM, Behringer RR et al. (2002) The novel zinc finger-containing transcription factor osterix is required for osteoblast differentiation and bone formation. Cell 108, 17-29. https://doi.org/10.1016/S0092-8674(01)00622-5
  10. Padgett RW, Wozney JM, Gelbart WM (1993) Human BMP sequences can confer normal dorsal-ventral patterning in the Drosophila embryo. Proc Natl Acad Sci USA 90, 2905-2909. https://doi.org/10.1073/pnas.90.7.2905
  11. Park SW, Kang SW, Goo TW, Kim SR, Paik SY (2012) Establishment of a Stable Cell Line Expressing Human BMP2/7-PTD for Efficient Osteogenic Induction. J Life Scienc 42, 456-465.
  12. Russell D, Sambrook J (2000) Molecular cloning; a laboratory manual. 3rd Ed, Cold Spring Harbor Laboratory Press, New York.
  13. Sampath TK, Rashka KE, Doctor JS, Tucker RF, Hoffmann FM (1993) Drosophila transforming growth factor beta superfamily proteins induce endochondral bone formation in mammals. Proc Natl Acad Sci U S A 90, 6004-6008. https://doi.org/10.1073/pnas.90.13.6004
  14. Sangadala S, Okada M, Liu Y, Viggeswarapu M, Titus L, Boden SD (2009) Engineering, cloning, and functional characterization of recombinant LIM mineralization protein-1 containing an N-terminal HIV-derived membrane transduction domain. Protein Expression Purifi 65, 165-173. https://doi.org/10.1016/j.pep.2009.01.006
  15. Wang X, Wenk E, Zhang X, Meinel L, Vunjak-Novakovic G, Kaplan DL (2009) Growth Factor Gradients via Microsphere Delivery in Biopolymer Scaffolds for Osteochondral Tissue Engineering. J Control Release 134, 81-90. https://doi.org/10.1016/j.jconrel.2008.10.021

Cited by

  1. Expression analysis ofdecapentaplegicgene in the silkworm,Bombyx mori vol.46, pp.6, 2016, https://doi.org/10.1111/1748-5967.12183
  2. Protein Expression of theBombyx mori DecapentaplegicGene using the Baculovirus Expression Vector System vol.45, pp.3, 2015, https://doi.org/10.4167/jbv.2015.45.3.256
  3. Expression of Recombinant Human Bone morphogenetic protein 2 (hBMP2) in Insect cells vol.34, pp.1, 2017, https://doi.org/10.7852/ijie.2017.34.1.1
  4. Enhancement of antimicrobial peptide genes expression in Cactus mutated Bombyx mori cells by CRISPR/Cas9 vol.37, pp.1, 2013, https://doi.org/10.7852/ijie.2018.37.1.21