Heat-induced apoptosis in human glioblastoma cell line A172.
OBJECTIVE
Hyperthermia has been clinically applied to some types of brain tumors. However, the detailed mechanisms of this growth inhibition are not clear. The effect of mild hyperthermia on cultured human glioblastoma cell line, A172, was studied.


METHODS
A172 cells were heat treated (43-44.5 degrees C) for 1 hour in the growing phase. Cell viability was assessed by trypan blue dye exclusion assay. The presence of apoptosis was determined by the morphological changes observed using phase contrast microscopy and nuclear changes observed using HOECHST 33342 stain. For the evaluation of cellular deoxyribonucleic acid fragmentation, the TUNEL method was used. The expression of p53 and bax proteins was evaluated by Western blot, and the bax messenger ribonucleic acid was detected by Northern blot.


RESULTS
Heat treatment induced cell death in time- and temperature-dependent manners. The nuclear staining with HOECHST 33342 demonstrated morphological changes consistent with apoptosis. The TUNEL stain also demonstrated damages in the deoxyribonucleic acid. These morphological changes were accompanied by the accumulation of p53 protein, bax protein, and messenger ribonucleic acid.


CONCLUSIONS
These results indicate that mild hyperthermia induces apoptosis in A172 glioblastoma cells.
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Department of Surgery, SLUCare, St. Louis, Missouri 63110-0250, USA.
Yoon
K W
KW
Kato
T
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Yamada
K
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D017209:Apoptosis / Q000502:physiology*; D001562:Benzimidazoles; D002470:Cell Survival / Q000502:physiology; D053938:DNA Fragmentation; D005456:Fluorescent Dyes; D005909:Glioblastoma / Q000473:pathology / Q000503:physiopathology*; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D008858:Microscopy, Phase-Contrast; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2; D012313:RNA / Q000096:biosynthesis; D012333:RNA, Messenger / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000187:drug effects; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein
C494826:BAX protein, human; D001562:Benzimidazoles; D005456:Fluorescent Dyes; D050884:HSP72 Heat-Shock Proteins; D006360:Heat-Shock Proteins; D011500:Protein Synthesis Inhibitors; D011518:Proto-Oncogene Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D012313:RNA; C017807:bisbenzimide ethoxide trihydrochloride