Role of mitochondria and calcium ions in tributyltin-induced gene regulatory pathways.
Tributyltin (TBT) salts are potent skin irritants both in humans and rodents. Data in the literature indicate mitochondria as target of TBT effects. Here, we investigate the early intracellular molecular events that follow TBT treatment and the relevance of calcium ions and mitochondria in gene-regulatory signaling pathways. Confluent HEL30 cells were treated with increasing doses of TBT (0-5 microM). At different times thereafter, the level of intracellular Ca2+, the cellular oxidative activity, nuclear factor-kappaB (NF-kappaB) activation, and IL-1alpha production were measured. TBT induced a dose-related increase of intracellular Ca2+ that reached the plateau 4 min following treatment. The increase of intracellular Ca2+ was followed by an increase in cellular oxidative activity as measured by DCFH oxidation (15 min) that preceded NF-kappaB activation (30 min) and IL-1alpha production (4 hr). All these events can be almost completely abrogated by BAPTA, an intracellular Ca2+ chelator. Furthermore, the modulation of cellular oxidative activity induced by TBT observed with rotenone, an inhibitor of the electron entry from complex I to ubiquinone, or after prolonged treatment with ethidium bromide, an inhibitor of mitochondrial DNA and RNA synthesis, indicates mitochondria as an important intracellular source of reactive oxygen species. These findings indicate the rise in intracellular Ca2+ as the starting event and indicate the role of mitochondria as the source of second messenger molecules essential for TBT-induced NF-kappaB activation and IL-1alpha production.
Corsini
E
E
Institute of Pharmacological Sciences, University of Milan, Italy.
Viviani
B
B
Marinovich
M
M
Galli
C L
CL
D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D002118:Calcium / Q000378:metabolism*; D002460:Cell Line; D002614:Chelating Agents / Q000494:pharmacology; D004272:DNA, Mitochondrial / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004533:Egtazic Acid / Q000031:analogs & derivatives / Q000494:pharmacology; D004797:Enzyme-Linked Immunosorbent Assay; D005452:Fluoresceins / Q000737:chemistry; D005456:Fluorescent Dyes / Q000737:chemistry; D005786:Gene Expression Regulation / Q000187:drug effects* / Q000235:genetics; D007375:Interleukin-1 / Q000096:biosynthesis / Q000378:metabolism; D015603:Keratinocytes / Q000166:cytology / Q000187:drug effects*; D051379:Mice; D008928:Mitochondria / Q000187:drug effects* / Q000378:metabolism; D008969:Molecular Sequence Data; D016328:NF-kappa B / Q000378:metabolism; D012313:RNA / Q000187:drug effects / Q000235:genetics / Q000378:metabolism; D000077278:RNA, Mitochondrial; D017382:Reactive Oxygen Species / Q000378:metabolism; D015290:Second Messenger Systems; D014220:Trialkyltin Compounds / Q000633:toxicity*
C096548:6-carboxy-2',7'-dichlorodihydrofluorescein diacetate bis(acetoxymethyl) ester; D002614:Chelating Agents; D004272:DNA, Mitochondrial; D005452:Fluoresceins; D005456:Fluorescent Dyes; D007375:Interleukin-1; D016328:NF-kappa B; D000077278:RNA, Mitochondrial; D017382:Reactive Oxygen Species; D014220:Trialkyltin Compounds; C011559:tributyltin; D004533:Egtazic Acid; D012313:RNA; C025603:1,2-bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid; D002118:Calcium