کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
10739273 | 1046869 | 2005 | 10 صفحه PDF | دانلود رایگان |
عنوان انگلیسی مقاله ISI
Role of Smac/DIABLO in hydrogen peroxide-induced apoptosis in C2C12 myogenic cells
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کلمات کلیدی
DIABLOPBSDMEMdirect IAP-binding protein with low pICytochrome oxidase subunit IIDTTIAPSCOX IISMACBSA - BSADulbecco's modified Eagle's medium - Medal of Eagle اصلاح شده DulbeccoROS - ROSHydrogen peroxide - آب اکسیژنهbovine serum albumin - آلبومین سرم گاوSmac/DIABLO - اسمک / DIABLOApoptosis - خزان یاختهایdithiothreitol - دیتیوتریتولphospate-buffered saline - فسفات بافر شورinhibitors of apoptosis proteins - مهارکننده های پروتئین آپوپتوزیسH2O2 - هیدروژن پراکسیدReactive oxygen species - گونههای فعال اکسیژن
موضوعات مرتبط
علوم زیستی و بیوفناوری
بیوشیمی، ژنتیک و زیست شناسی مولکولی
سالمندی
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چکیده انگلیسی
Smac/DIABLO was recently identified as a protein released from mitochondria in response to apoptotic stimuli which promotes apoptosis by antagonizing inhibitors of apoptosis proteins. Furthermore, Smac/DIABLO plays an important regulatory role in the sensitization of cancer cells to both immune-and drug-induced apoptosis. However, little is known about the role of Smac/DIABLO in hydrogen peroxide (H2O2)-induced apoptosis of C2C12 myogenic cells. In this study, Hoechst 33258 staining was used to examine cell morphological changes and to quantitate apoptotic nuclei. DNA fragmentation was observed by agarose gel electrophoresis. Intracellular translocation of Smac/DIABLO from mitochondria to the cytoplasm was observed by Western blotting. Activities of caspase-3 and caspase-9 were assayed by colorimetry and Western blotting. Full-length Smac/DIABLO cDNA and antisense phosphorothioate oligonucleotides against Smac/DIABLO were transiently transfected into C2C12 myogenic cells and Smac/DIABLO protein levels were analyzed by Western blotting. The results showed that: (1) H2O2 (0.5 mmol/L) resulted in a marked release of Smac/DIABLO from mitochondria to cytoplasm 1 h after treatment, activation of caspase-3 and caspase-9 4 h after treatment, and specific morphological changes of apoptosis 24 h after treatment; (2) overexpression of Smac/DIABLO in C2C12 cells significantly enhanced H2O2-induced apoptosis and the activation of caspase-3 and caspase-9 (PÂ <Â 0.05). (3) Antisense phosphorothioate oligonucleotides against Smac/DIABLO markedly inhibited de novo synthesis of Smac/DIABLO and this effect was accompanied by decreased apoptosis and activation of caspase-3 and caspase-9 induced by H2O2 (PÂ <Â 0.05). These data demonstrate that H2O2 could result in apoptosis of C2C12 myogenic cells, and that release of Smac/DIABLO from mitochondria to cytoplasm and the subsequent activation of caspase-9 and caspase-3 played important roles in H2O2-induced apoptosis in C2C12 myogenic cells.
ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Free Radical Biology and Medicine - Volume 39, Issue 5, 1 September 2005, Pages 658-667
Journal: Free Radical Biology and Medicine - Volume 39, Issue 5, 1 September 2005, Pages 658-667
نویسندگان
Bimei Jiang, Weimin Xiao, Yongzhong Shi, Meidong Liu, Xianzhong Xiao,