Identification of to Hexapeptides that Render C2 Myoblasts the Resistant Menadione-induced Cell Death

  • Hwang, Sung-Ho (Department of Biological Science, College of Natural Sciences, Ajou University) ;
  • Kim, Min-Jeong (Department of Biological Science, College of Natural Sciences, Ajou University) ;
  • Lim, Jeong-A (Department of Biological Science, College of Natural Sciences, Ajou University) ;
  • Woo, Joo-Hong (Department of Biological Science, College of Natural Sciences, Ajou University) ;
  • Kim, Hye-Sun (Department of Biological Science, College of Natural Sciences, Ajou University)
  • Published : 2008.03.31

Abstract

Menadione induced cell death in cultured C2 myoblasts. By screening synthetic peptide libraries composed of random sequence of hexapeptides, we identified the hexa-peptides pool of(Ala/Ile)-(Ile/Met)-Val-Ile-Asp-(Met/Ser)-$NH_2$ that protected the myoblasts against menadioneinduced cell death. Pre-incubation with the hexapeptide pool reduced the number of cells detached from culture dish substrate and increased the ratio of relative viability against menadione. In addition, the peptides strongly increased the expression of Bcl-2, an anti-apoptotic protein. These results suggest that the hexapeptides might enhance the resistance to cell death against menadione by increasing the expression of Bcl-2.

Keywords

References

  1. Basset O, Boittin FX, Cognard C, Constantin B, and Ruegg UT (2006) Bcl-2 overexpression prevents calcium overload and subsequent apoptosis in dystrophic myotubes. Biochem J 395:267-276 https://doi.org/10.1042/BJ20051265
  2. Burlacu A (2003) Regulation of apoptosis by Bcl-2 family proteins. J Cell Mol Med 7:249-257 https://doi.org/10.1111/j.1582-4934.2003.tb00225.x
  3. Chen YR, Wang W, Kong AN, and Tan TH (1998) Molecular mechanisms of c-Jun N-terminal kinase-mediated apoptosis induced by anticarcinogenic isothiocyanates. J Biol Chem 273:1769-1775 https://doi.org/10.1074/jbc.273.3.1769
  4. Chiou TJ, Chu ST, and Tzeng WF (2003) Protection of cells from menadioneinduced apoptosis by inhibition of lipid peroxidation. Toxicology 191:77-88 https://doi.org/10.1016/S0300-483X(03)00189-6
  5. Dominov JA, Kravetz AJ, Ardelt M, Kostek CA, Beermann ML, and Miller JB (2005) Muscle-specific BCL2 expression ameliorates muscle disease in laminin {alpha}2-deficient, but not in dystrophin-deficient, mice. Hum Mol Genet 14:1029-1040 https://doi.org/10.1093/hmg/ddi095
  6. Girgenrath M, Dominov JA, Kostek CA, and Miller JB (2004) Inhibition of apoptosis improves outcome in a model of congenital muscular dystrophy. J Clin Invest 114:1635-1639 https://doi.org/10.1172/JCI22928
  7. Haddad JJ (2004) On the antioxidant mechanisms of Bcl-2: a retrospective of NF-kappaB signaling and oxidative stress. Biochem Biophys Res Commun 322:355-363 https://doi.org/10.1016/j.bbrc.2004.07.138
  8. Houghten RA, Pinilla C, Appel JR, Blondelle SE, Dooley CT, Eichler J, Nefzi A, and Ostresh JM (1999) Mixture-based synthetic combinatorial libraries. J Med Chem 42:3743-3778 https://doi.org/10.1021/jm990174v
  9. Houghten RA, Pinilla C, Blondelle SE, Appel JR, Dooley CT, and Cuervo JH (1991) Generation and use of synthetic peptide combinatorial libraries for basic research and drug discovery. Nature 354:84-86 https://doi.org/10.1038/354084a0
  10. Kefaloyianni E, Gaitanaki C, and Beis I (2006) ERK1/2 and p38-MAPK signalling pathways, through MSK1, are involved in NF-kappaB transactivation during oxidative stress in skeletal myoblasts. Cell Signal 18:2238-2251 https://doi.org/10.1016/j.cellsig.2006.05.004
  11. McArdle F, Pattwell DM, Vasilaki A, McArdle A, and Jackson MJ (2005) Intracellular generation of reactive oxygen species by contracting skeletal muscle cells. Free Radic Biol Med 39:651-657 https://doi.org/10.1016/j.freeradbiomed.2005.04.010
  12. Metrailler-Ruchonnet I, Pagano A, Carnesecchi S, Ody C, Donati Y, Barazzone and Argiroffo C (2007) Bcl-2 protects against hyperoxia-induced apoptosis through inhibition of the mitochondria-dependent pathway. Free Radic Biol Med 42:1062-1074 https://doi.org/10.1016/j.freeradbiomed.2007.01.008
  13. Monks TJ, Hanzlik RP, Cohen GM, Ross D, and Graham DG (1992) Quinone chemistry and toxicity. Toxicol Appl Pharmacol 112:2-16 https://doi.org/10.1016/0041-008X(92)90273-U
  14. Niwa K, Inanami O, Yamamori T, Ohta T, Hamasu T, and Kuwabara M (2003) Redox regulation of PI3K/Akt and p53 in bovine aortic endothelial cells exposed to hydrogen peroxide. Antioxid & Redox Signaling 5:713-722 https://doi.org/10.1089/152308603770380016
  15. Park JY, Kim IJ, Lee MH, Seo JK, Suh PG, Cho BY, Ryu SH, and Chae CB (1997) Identification of the peptides that inhibit the stimulation of thyrotropin receptor by Graves' immunoglobulin G from peptide libraries. Endocrinology 138:617-626 https://doi.org/10.1210/en.138.2.617
  16. Pinilla C, Appel JR, Blanc P, and Houghten RA (1992) Rapid identification of high affinity peptide ligands using positional scanning synthetic peptide combinatorial libraries. Biotechniques 13:901-905
  17. Pinilla C, Appel JR, Borras E, and Houghten RA (2003) Advances in the use of synthetic combinatorial chemistry: mixture-based libraries. Nat Med 9:118-122 https://doi.org/10.1038/nm0103-118
  18. Rando TA, Disatnik MH, Yu Y, and Franco A (1998) Muscle cells from mdx mice have an increased susceptibility to oxidative stress. Neuromuscul Disord 8:14-21 https://doi.org/10.1016/S0960-8966(97)00124-7
  19. Rudolf E, and Cervinka M (2006) Cytoskeletal changes in nonapoptotic cell death. Acta Medica (Hradec Kralove) 49:123-128
  20. Shimada K, Nakamura M, Ishida E, Kishi M, and Konishi N (2003) Roles of p38- and c-jun NH2-terminal kinasemediated pathways in 2-methoxyestradiol-induced p53 induction and apoptosis. Carcinogenesis 24:1067-1075 https://doi.org/10.1093/carcin/bgg058
  21. Sun JS, Tsuang YH, Huang WC, Chen LT, Hang YS, and Lu FJ (1997) Menadione-induced cytotoxicity to rat osteoblasts. Cell Mol Life Sci 53:967-976 https://doi.org/10.1007/s000180050118
  22. Wagstaff KM, and Jans DA (2006) Protein transduction: cell penetrating peptides and their therapeutic applications. Curr Med Chem 13:1371-1387 https://doi.org/10.2174/092986706776872871
  23. Woo JH, and Kim HS (2006) Phosphorylation of eukaryotic elongation factor 2 can be regulated by phosphoinositide 3- kinase in the early stages of myoblast differentiation. Mol Cells 21:294-301