Establishment And Application Of HEK293 Cell Lines Stably Expressing μ Opioid Receptor Or δ Opioid Receptor

μopioid receptors and 8 opioid receptors display widespread locatization in the central and peripheral nervous systems. Both of them participate in a broad range of biological processes, including nociception, cardiovascular, respiratory, etc.μreceptor agonists such as morphine and heroin are among the most effective analgesics known. However, their efficacy is accompanied by burdensome side effects. While 8 receptor agonists with weaker analgesic effects can produce less side effects thanμreceptor agonists. In order to research the affinity and activity of opioid peptides and receptors on the cellular level, we established HEK293 cells which could stably expressμreceptors orδreceptors. First, we constructed two recombinant plasmids pcDNA3.1-FLAG-MOP and pcDNA3.1-MYC-DOP. Second, HEK293 cells were transfected with the plasmid pcDNA3.1-FLAG-MOP or pcDNA3.1-MYC-DOP by using Lipofectamine 2000. Then, we selected the positive clones which could stably express one kind of the two receptors, and the positive clones were confirmed by RT-PCR, intracellular Ca2+ release and Western blotting. We used the two cell models to research the activity of 8 endomorphin analogs in radioligand competitive binding assay, cAMP and ERK 1/2 signaling pathways. The two cell models can be used to the study of cell signaling mediated byμreceptors orδreceptors.