Opioid receptor gene: cytokine response element and the effect of cytokines

Previous studies demonstrated that cytokines regulate opioid and opioid receptor gene expression in neuronal and immune cells. The gene sequence analysis of opioid receptors revealed that mu-, delta- and kappa-opioid receptor promoter regions contain potential cytokine response elements (NF-IL6 binding sites). It was postulated that the response elements present in opioid receptor promoter regions may have a role in the cytokine effects on opioid receptor gene expression through cis-trans interaction. The present study investigated whether cytokines have an effect on opioid receptor gene expression by cytokine-induced transcription factor, NF-IL6, using a number of immune cell lines which respond to cytokines. Further investigation was made to determine whether the potential cytokine response element DNA sequences on opioid receptor promoter region have functional significance which may be affected by the DNA context of the opioid receptor promoter in immune cell lines. Tandem repeats of conserved cytokine response elements of IL-6 gene fused to a heteropromoter SV40 were utilized as a positive control and expressed 2-fold increased promoter activity after cytokine stimulation. Transient transfection studies in time courses (24-72 h) and different dose treatments (100-500 U/ml for IL-6 and 50-200 U/ml for IL-1 alpha+beta) were also carried out to investigate the possibility that the upregulated gene expression may be transient or cytokine-dose-dependent. Our data demonstrated that there was no significant cytokine-stimulated opioid receptor gene expression in immune cells tested. In addition, the cytokine response elements (NF-IL6 binding sites) in opioid receptor genes are not functional. These results contradict the previous reports in which cytokines modulated the expression of opioid and opioid receptors in neuronal and immune cells. The possible reasons regarding the different results were discussed.