Mechanism of heme oxygenase-1 gene activation by cadmium in MCF-7 mammary epithelial cells - Role of p38 kinase and Nrf2 transcription factor

The mouse heme oxygenase-1 (HO-1) gene, ho-1, contains two inducible enhanc ers, E1 and E2. Of several cell Lines tested, induction of an E1/luciferase fusion construct, pE1-luc, by CdCL2 is most pronounced in MCF-7 cells. In these cells, El, but not E2, is necessary and sufficient for ho-1 gene acti vation. Exposure of MCF-7 cells to 10 mu M CdCl2 stimulates phosphorylation of ERK, JNK, and p38 mitogen-activated protein kinases, implicating one or more of these signaling pathways in ho-1 gene induction. SB203580, an inhi bitor of p38, diminishes cadmium-stimulated pE1-luc expression and HO-1 mRN A levels by up to 70-80%. PD098059, an ERK pathway inhibitor, does not affe ct HO-1 mRNA induction at the highest concentration (40 mu M) tested. Simil arly, coexpression of a dominant-negative mutant of p38 alpha, but not of E RK1, ERK2, JNK1, or JNK2, reduces basal and cadmium-induced pE1-luc activit y. E1 contains binding sites for the activator protein-1 (Fos/Jun), Cap'n'C ollar/basic leucine zipper (CNC-bZIP), and CCAAT/enhancer-binding protein ( C/EBP) families of transcription factors. A dominant-negative mutant of Nrf 2 (a CNC-bZIP member), but not of c-Jun or C/EBP beta, inhibits pE1-luc act ivation by cadmium. Induction of the endogenous ho-1 gene is also inhibited by the Nrf2 mutant. Flutations of E1 that inhibit cadmium inducibility als o suppress the trans-activation and DNA binding activities of Nrf2, and SB2 03580, but not PD098059, attenuates Nrf2-mediated trans-activation of pE1-l uc. Taken together, these results indicate that cadmium induces ho-1 gene e xpression via sequential activation of the p38 kinase pathway and Nrf2.