Induction of heat shock protein 72 synthesis by endogenous tumor necrosis factor via enhancement of the heat shock element-binding activity of heat shock factor 1.
Endogenous tumor necrosis factor (enTNF) acts as a resistance factor against cytotoxicity caused by heat by inducing manganous superoxide dismutase (MnSOD), thereby scavenging reactive oxygen free radicals. On the other hand, it is also well known that heat shock proteins (HSP) which are induced by heat stress behave as cytoprotective factor against this stress. However, the relationship of these two resistance factors is not elucidated yet. In the present study, we therefore proposed the possibility that enTNF enhances HSP72 expression. Heat-sensitive L-M (mouse tumorigenic fibroblast) cells, which normally do not express enTNF, were transfected with a nonsecretory-type human TNF-alpha expression vector to produce enTNF. Stable transfectants showed resistance to heat treatment and an increase of HSP72 expression. Conversely, when HeLa (human uterine cervical cancer) cells, which normally produce an appreciable amount of enTNF, were transfected with an antisense TNF-alpha mRNA expression vector to inhibit enTNF synthesis, their heat sensitivity was enhanced and HSP72 expression was reduced by half. Although enTNF caused no difference in the level of heat shock factor (HSF) 1 in these cells, enTNF expression correlated well with the binding activity of HSF-1 to a 32P-labeled synthetic oligonucleotide containing the human heat shock element (HSE). These results indicate that enTNF participates not only in intrinsic resistance against heat via induction of MnSOD but also via enhancement of the HSE-binding activity of HSF 1 followed by augmentation of HSP72 expression.
Watanabe
N
N
Department of Laboratory Diagnosis, Sapporo Medical University, School of Medicine, Japan. watanabn@sapmed.ac.jp
Tsuji
N
N
Akiyama
S
S
Sasaki
H
H
Okamoto
T
T
Kobayashi
D
D
Sato
T
T
Hagino
T
T
Yamauchi
N
N
Niitsu
Y
Y
Nakai
A
A
Nagata
K
K
D000818:Animals; D016375:Antisense Elements (Genetics) / Q000502:physiology; D004268:DNA-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000502:physiology*; D005786:Gene Expression Regulation / Q000276:immunology; D005822:Genetic Vectors / Q000378:metabolism; D050884:HSP72 Heat-Shock Proteins; D006367:HeLa Cells; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D006358:Hot Temperature; D006801:Humans; D007739:L Cells; D051379:Mice; D014157:Transcription Factors; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis / Q000235:genetics / Q000502:physiology*
D016375:Antisense Elements (Genetics); D004268:DNA-Binding Proteins; D050884:HSP72 Heat-Shock Proteins; D000076249:Heat Shock Transcription Factors; D006360:Heat-Shock Proteins; D014157:Transcription Factors; D014409:Tumor Necrosis Factor-alpha