Recombinant heat shock protein 27 (HSP27/HSPB1) protects against cadmium-induced oxidative stress and toxicity in human cervical cancer cells.
Cadmium (Cd) is a carcinogen with several well-described toxicological effects in humans, but its molecular mechanisms are still not fully understood. Overexpression of heat shock protein 27 (HSP27/HSPB1)-a multifunctional protein chaperone-has been shown to protect cells from oxidative damage and apoptosis triggered by Cd exposure. The aims of this work were to investigate the potential use of extracellular recombinant HSP27 to prevent/counteract Cd-induced cellular toxicity and to evaluate if peroxynitrite was involved in the development of Cd-induced toxicity. Here, we report that the harmful effects of Cd correlated with changes in oxidative stress markers: upregulation of reactive oxygen species, reduction in nitric oxide (NO) bioavailability, increment in lipid peroxidation, peroxynitrite (PN), and protein nitration; intracellular HSP27 was reduced. Treatments with Cd (100 μM) for 24 h or with the peroxynitrite donor, SIN-1, decreased HSP27 levels (~50%), suggesting that PN formation is responsible for the reduction of HSP27. Pre-treatments of the cells either with Nω-nitro-L-arginine methyl ester hydrochloride (L-NAME) (a pharmacological inhibitor of NO synthase) or with recombinant HSP27 (rHSP27) attenuated the disruption of the cellular metabolism induced by Cd, increasing in a 55 and 52%, respectively, the cell viability measured by CCK-8. Cd induced necrotic cell death pathways, although apoptosis was also activated; pre-treatment with L-NAME or rHSP27 mitigated cell death. Our findings show for the first time a direct relationship between Cd-induced toxicity and PN production and a role for rHSP27 as a potential therapeutic agent that may counteract Cd toxicity.
Oncology Laboratory at the Institute of Experimental Medicine and Biology of Cuyo (IMBECU), CCT CONICET Mendoza, Av. Ruiz Leal S/N Parque Gral. San Martin, CP 5500, Mendoza, CP, Argentina.|Libin Cardiovascular Institute of Alberta, Cumming School of Medicine, Foothills Medical Centre, University of Calgary, Room C823, 1403-29th Street NW, Calgary, AB, T2N 2T9, Canada.
Daiana G
DG
Alvarez-Olmedo
Department of Biochemistry and Biological Sciences, Faculty of Chemistry, Biochemistry, and Pharmacy, National University of San Luis, IMIBIO-SL, CONICET, San Luis, Argentina.
Veronica S
VS
Biaggio
Libin Cardiovascular Institute of Alberta, Cumming School of Medicine, Foothills Medical Centre, University of Calgary, Room C823, 1403-29th Street NW, Calgary, AB, T2N 2T9, Canada.
Geremy A
GA
Koumbadinga
Department of Biochemistry and Biological Sciences, Faculty of Chemistry, Biochemistry, and Pharmacy, National University of San Luis, IMIBIO-SL, CONICET, San Luis, Argentina.
Nidia N
NN
Gómez
Libin Cardiovascular Institute of Alberta, Cumming School of Medicine, Foothills Medical Centre, University of Calgary, Room C823, 1403-29th Street NW, Calgary, AB, T2N 2T9, Canada.
Chunhua
C
Shi
Oncology Laboratory at the Institute of Experimental Medicine and Biology of Cuyo (IMBECU), CCT CONICET Mendoza, Av. Ruiz Leal S/N Parque Gral. San Martin, CP 5500, Mendoza, CP, Argentina.
Daniel R
DR
Ciocca
Libin Cardiovascular Institute of Alberta, Cumming School of Medicine, Foothills Medical Centre, University of Calgary, Room C823, 1403-29th Street NW, Calgary, AB, T2N 2T9, Canada.
Zarah
Z
Batulan
Oncology Laboratory at the Institute of Experimental Medicine and Biology of Cuyo (IMBECU), CCT CONICET Mendoza, Av. Ruiz Leal S/N Parque Gral. San Martin, CP 5500, Mendoza, CP, Argentina. mfanelli@mendoza-conicet.gob.ar.
Mariel A
MA
Fanelli
Libin Cardiovascular Institute of Alberta, Cumming School of Medicine, Foothills Medical Centre, University of Calgary, Room C823, 1403-29th Street NW, Calgary, AB, T2N 2T9, Canada. ermobrie@ucalgary.ca.
Edward R
ER
O'Brien
D017209:Apoptosis / Q000187:drug effects; D002104:Cadmium / Q000633:toxicity*; D002465:Cell Movement / Q000187:drug effects; D005260:Female; D005456:Fluorescent Dyes / Q000737:chemistry; D055551:HSP27 Heat-Shock Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D006367:HeLa Cells; D006801:Humans; D008856:Microscopy, Fluorescence; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism; D018384:Oxidative Stress / Q000187:drug effects*; D030421:Peroxynitrous Acid / Q000032:analysis / Q000378:metabolism; D017382:Reactive Oxygen Species / Q000378:metabolism; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000302:isolation & purification / Q000494:pharmacology; D015854:Up-Regulation / Q000187:drug effects; D002583:Uterine Cervical Neoplasms / Q000378:metabolism / Q000473:pathology
D005456:Fluorescent Dyes; D055551:HSP27 Heat-Shock Proteins; D017382:Reactive Oxygen Species; D011994:Recombinant Proteins; D002104:Cadmium; D030421:Peroxynitrous Acid; D009569:Nitric Oxide; D019331:NG-Nitroarginine Methyl Ester
Cytoprotection; Heat shock protein 27 (HSP27); Oxidative stress; Signaling cadmium toxicity