Constitutive expression of human hsp27, Drosophila hsp27, or human alpha B-crystallin confers resistance to TNF- and oxidative stress-induced cytotoxicity in stably transfected murine L929 fibroblasts.
Hyperthermia and other forms of stress that induce and/or stimulate heat shock or stress protein (hsp) expression enhance the cellular resistance to TNF-alpha. One of the stress proteins, hsp70, has been shown to participate in the molecular mechanisms that regulate this phenomenon. Here we have tested the capability of small hsps from different species to protect against this cytokine in the TNF-sensitive L929 fibrosarcoma cells. The genes that encode human hsp27, Drosophila hsp27, and human alpha B-crystallin were placed under the control of the constitutive SV40 early promoter and were stably introduced into L929 cells. We observed that all clones that constitutively expressed the exogenous small hsps exhibited a strong protection against TNF-mediated killing, which was proportional to the level of the expressed proteins. This phenomenon did not correlate with altered binding of TNF-alpha to its receptors, and no protection was observed as a consequence of the transfection or selection procedures. In addition, the overexpression of the exogenous small hsps did not modify the level of the endogenous stress proteins in the transfected clones. Remarkably, the small hsps also induced a protection against oxidative stresses generated by either hydrogen peroxide or menadione. In L929 cells, the killing induced by TNF-alpha and oxidative stress is thought to occur through the accumulation of intracellular reactive oxygen intermediates. Hence, our data suggest that the small hsps from different species share the property to protect L929 cells against the deleterious effects of reactive oxygen intermediates generated by either TNF-alpha or oxidative stress.
Mehlen
P
P
Laboratory of Cellular Stress, Claude Bernard University, Lyon, Villeurbanne, France.
Preville
X
X
Chareyron
P
P
Briolay
J
J
Klemenz
R
R
Arrigo
A P
AP
D000818:Animals; D003459:Crystallins / Q000096:biosynthesis* / Q000235:genetics; D004331:Drosophila melanogaster / Q000235:genetics; D004351:Drug Resistance; D015967:Gene Expression Regulation, Viral; D005813:Genes, Synthetic; D006360:Heat-Shock Proteins / Q000096:biosynthesis* / Q000235:genetics; D006801:Humans; D006861:Hydrogen Peroxide / Q000494:pharmacology; D007739:L Cells / Q000187:drug effects / Q000378:metabolism; D051379:Mice; D018384:Oxidative Stress; D011401:Promoter Regions, Genetic; D017382:Reactive Oxygen Species / Q000633:toxicity; D018124:Receptors, Tumor Necrosis Factor / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis*; D011994:Recombinant Proteins / Q000494:pharmacology; D013539:Simian virus 40 / Q000235:genetics; D013045:Species Specificity; D013312:Stress, Physiological / Q000378:metabolism*; D014162:Transfection; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology*; D014812:Vitamin K / Q000494:pharmacology
D003459:Crystallins; D006360:Heat-Shock Proteins; D017382:Reactive Oxygen Species; D018124:Receptors, Tumor Necrosis Factor; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D014409:Tumor Necrosis Factor-alpha; D014812:Vitamin K; D006861:Hydrogen Peroxide