Nitric oxide induces apoptosis in mouse C2C12 myoblast cells.
To investigate whether nitric oxide (NO) induces apoptosis in myoblast cells, the effect of the sodium nitroprusside (SNP), NO donor, on the apoptosis of mouse C2C12 myoblast cells was examined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, flow cytometry, 4,6-diamidino-2-phenylindole (DAPI) staining, terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL) assay, DNA fragmentation assay, reverse transcription-polymerase chain reaction (RT-PCR), Western blot analysis, and caspase-3 enzyme assay. Mouse C2C12 myoblast cells treated with SNP exhibited several apoptotic features. SNP increased p53 expression and bax expression. SNP also enhanced caspase-3 enzyme activity. The data show that NO may induce apoptotic cell death in myoblast cells through the activation of p53-, bax-, and caspase-dependent intracellular death-related pathways.
Lee
Myoung-Hwa
MH
Department of Physiology, College of Medicine, Kyung Hee University, Seoul, Korea.
Jang
Mi-Hyeon
MH
Kim
Eun-Kyung
EK
Han
Sang-Wan
SW
Cho
Seong-Yeon
SY
Kim
Chang-Ju
CJ
D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D015153:Blotting, Western; D053148:Caspase 3; D020169:Caspases / Q000378:metabolism; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D053938:DNA Fragmentation / Q000187:drug effects; D004305:Dose-Response Relationship, Drug; D004792:Enzyme Precursors / Q000378:metabolism; D005434:Flow Cytometry; D005456:Fluorescent Dyes; D020287:In Situ Nick-End Labeling; D007211:Indoles; D051379:Mice; D008856:Microscopy, Fluorescence; D032446:Myoblasts / Q000187:drug effects*; D009569:Nitric Oxide / Q000494:pharmacology*; D020030:Nitric Oxide Donors; D009599:Nitroprusside / Q000494:pharmacology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013997:Time Factors; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein
C494827:Bax protein, mouse; D004792:Enzyme Precursors; D005456:Fluorescent Dyes; D007211:Indoles; D020030:Nitric Oxide Donors; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D009599:Nitroprusside; D009569:Nitric Oxide; C007293:DAPI; C505409:Casp3 protein, mouse; D053148:Caspase 3; D020169:Caspases