Role of Smac/DIABLO in H2O2-induced Apoptosis in C2C12 Myogenic Cells

Bi Mei Jiang, Wei Min Xiao, Yong Zhong Shi, Mei Dong Liu, Dao Lin Tang, Xian Zhong Xiao

Research output: Contribution to journalArticle

Abstract

In order to explore the role of Smac/DIABLO in H2O 2-induced apoptosis of C2C12 myogenic cells, Hoechst 33258 staining was used to examine cell morphological changes and to calculate percentage of apoptotic nuclei. DNA ladder pattern on agarose gel electrophoresis was used to observe DNA fragmentation. The release of Smac/DIABLO from mitochondria to cytoplasm was observed by Western blotting. Activities of Caspase-3, Caspase-9 were assayed with Caspase Colorimetric Assay Kit and Western blotting. Full length Smac/DIABLO gene was transiently transfected in C2C12 myogenic cells by lipofectamine and then protein levels of Smac/DIABLO were analysed by Western blotting. The results showed that; (1) H2O2 (O. 5 mmol/L) resulted in a marked release of Smac/DIABLO from mitochondrial to cytoplasm l h after treatment, activation of Caspase-3, Caspase-9 4 h after treatment and specific morphological changes of apoptosis 24 h after treatment; (2) Smac/DIABLO overexpression significantly enhanced H2O2 induced apoptosis in C2C12 myogenic cells as shown by specific DNA ladder pattern in agarose gel electrophoresis, increase of percentage of apoptotic nuclei and marked activation of Caspase-3, Caspase-9. These data suggested that H2O2 could result in apoptosis of C 2C12 myogenic cells, and that release of Smac/DIABLO from mitochondrial to cytoplasm and the subsequent activation of Caspase-9 and Caspase-3 played important roles in H2O2-induced apoptosis in C2C12 myogenic cells.

Original languageEnglish (US)
Pages (from-to)923-929
Number of pages7
JournalProgress in Biochemistry and Biophysics
Volume30
Issue number6
Publication statusPublished - Dec 1 2003
Externally publishedYes

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Keywords

  • Apoptosis
  • CC myogenic cells
  • Hydrogen peroxide
  • Smac/DIABLO

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry

Cite this

Jiang, B. M., Xiao, W. M., Shi, Y. Z., Liu, M. D., Tang, D. L., & Xiao, X. Z. (2003). Role of Smac/DIABLO in H2O2-induced Apoptosis in C2C12 Myogenic Cells. Progress in Biochemistry and Biophysics, 30(6), 923-929.