Cd(2+)-induced c-myc mRNA accumulation in NRK-49F cells is blocked by the protein kinase inhibitor H7 but not by HA1004, indicating that protein kinase C is a mediator of the response.
Cd2+ is a toxic cation that, at sublethal and marginally lethal levels, modifies cell growth and metabolism. Cd2+ exposure of NRK-49F cells results in inhibition of early EGF-induced DNA synthesis, but induction of delayed DNA synthesis; in stimulation of anchorage independent growth; in accumulation of specific oncogene mRNAs; and in an hypertrophic response. Determining whether specific signal transduction pathways (STPs) are involved in specific gene deregulation by cadmium in NRK-49F cells is important to defining possible mechanisms by which Cd2+ elicits these physiological responses. In this study it is shown that Cd2+ induces delayed myc (8-10 h) and jun (12 h) mRNA accumulation, as well as both early (0.5-1 h) and late (12 h) fos but not TGF beta mRNA accumulation. The times of appearance of Cd(2+)-induced c-fos, c-myc and c-jun expression are dose dependent. The Cd2+ induced accumulation of these specific mRNAs is insensitive to cycloheximide and therefore not due to preinduction of TGF beta or other gene-activating growth factors, but rather to direct induction of oncogene expression and/or mRNA stabilization. Accumulation of c-myc mRNA is shown further to be inhibited by the protein kinase inhibitor H7 but not HA1004, indicating a role for one or more protein kinases C in the STPs by which Cd2+ induces oncogene expression. Thapsigargin, a compound which stimulates increased cytosolic [Ca2+], induces c-myc expression also by an H7 sensitive, HA1004 insensitive pathway. These results suggest that Cd2+ acts through one or more defined signal transduction pathways involving specific protein kinases C to induce the accumulation of c-fos, c-myc and c-jun messenger RNAs.
Department of Zoology and Genetics, Iowa State University, Ames 50011.
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D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine / Q000031:analogs & derivatives*; D000818:Animals; D015152:Blotting, Northern; D002104:Cadmium / Q000494:pharmacology / Q000633:toxicity*; D002460:Cell Line; D015342:DNA Probes; D015870:Gene Expression / Q000187:drug effects; D016762:Genes, fos / Q000187:drug effects / Q000235:genetics; D016758:Genes, jun / Q000187:drug effects / Q000235:genetics; D016259:Genes, myc / Q000187:drug effects* / Q000235:genetics; D007546:Isoquinolines / Q000494:pharmacology*; D010879:Piperazines / Q000494:pharmacology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger / Q000378:metabolism*; D051381:Rats; D013449:Sulfonamides
D015342:DNA Probes; D007546:Isoquinolines; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; D013449:Sulfonamides; D002104:Cadmium; C080501:1-(5-isoquinolinylsulfonyl)-3-methylpiperazine; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; C043204:N-(2-guanidinoethyl)-5-isoquinolinesulfonamide; D011493:Protein Kinase C