Dual effect of 1 alpha,25-dihydroxyvitamin D3 on hsp28 and PKC beta gene expression in phorbol ester-resistant human myeloid HL-525 leukemic cells.

We investigated the effect of 1 alpha-25-dihydroxyvitamin D3 [1,25-(OH)2D3] on the expression of the 28-kDa heat shock protein gene (hsp28) and the protein kinase C beta gene (PKC beta) in the human myeloid HL-60 leukemic cell variant HL-525, which is resistant to phorbol ester-induced macrophage differentiation. Northern and western blot analysis showed little or no hsp28 gene expression in the HL-60 cell variant, HL-205, which is susceptible to such differentiation, while a relatively high basal level of hsp28 gene expression was observed in the HL-525 cells. However, both cell lines demonstrated heat shock-induced expression of this gene. During treatment with 50-300 nM 1,25-(OH)2D3, a marked reduction of hsp28 gene expression along with an induction of PKC beta gene expression was observed in HL-525 cells. A gel mobility-shift assay demonstrated that the 1,25-(OH)2D3-induced alteration of hsp28 gene expression was associated with decreased binding activity to the vitamin D3 receptor-vitamin D3 response element (VDR-VDRE), whereas binding to the heat shock transcription factor-heat shock element (HSF-HSE) was not altered. Our results suggest that the dual effect of 1,25-(OH)2D3 on hsp28 and PKC beta gene expression is due to the different sequence composition of the vitamin D response element in the promoter region as well as an accessory factor for each gene or that 1,25-(OH)2D3 increases PKC beta gene expression, which, in turn, negatively regulates the expression of the hsp28 gene or vice versa.