title: Different role of cAMP pathway on the human mast cells HMC-1(560) and HMC-1(560,816) activation.
abstract: HMC-1 are inflammatory cells that release vasoactive substances such as histamine. These cells have the c-kit receptor permanently activated in the membrane due to mutations in the proto-oncogene c-kit: Val-560 → Gly and Asp-816 → Val. Thus, there are two known cellular lines: HMC-1(560) and HMC-1(560,816) . These mutations are involved in a disease called mastocitosys. In the present paper both lines were used to study the influence of cAMP/PKA/PDEs pathway on the histamine release and Ca(2+) signaling since this pathway is often involved in these process. For this, the cells were preincubated with cAMP/PKA/PDEs modulators such as dibutyryl cAMP (dbcAMP), forskolin, H89, rolipram, IBMX, or imidazole and then stimulated with ionomycin. When cells were stimulated with agents that increase cAMP levels, the histamine release was not modified in HMC-1(560) but decreased in HMC-1(560,816) cells. The same happened when PKA was blocked. Furthermore, PDEs role on histamine release was independent of cAMP in HMC-1(560) cells and possibly also in HMC-1(560,816) cells. By contrast, the modulation of PKA and PDEs together changed the response in both cellular lines, therefore a relationship between them was suggested. All these modulatory effects on histamine release are Ca(2+) -independent. On the other hand, the effect of c-kit modulation on the cAMP/PKA/PDEs pathway was also checked. This receptor was blocked with STI571 (imatinib) and BMS-354825 (dasatinib), but only the last one caused a decrease in the cellular response to ionomycin. This article demonstrates for the first time than the cAMP/PKA/PDEs pathway is involved in the activation of HMC-1(560) and HMC-1(560,816) cells.
authors.affiliation: Department of Farmacología, Facultad de Veterinaria, 27002, Lugo, Spain.
authors.forename: Olalla
authors.initials: O
authors.lastname: Barreiro-Costa
authors.forename: Araceli
authors.initials: A
authors.lastname: Tobío
authors.forename: Amparo
authors.initials: A
authors.lastname: Alfonso
authors.forename: Luis M
authors.initials: LM
authors.lastname: Botana
mesh_terms: D020013:Calcium Signaling / Q000187:drug effects / Q000235:genetics; D002460:Cell Line; D000242:Cyclic AMP / Q000378:metabolism*; D006801:Humans; D008407:Mast Cells / Q000378:metabolism*; D011493:Protein Kinase C / Q000378:metabolism; D000090063:Proto-Oncogene Mas; D019009:Proto-Oncogene Proteins c-kit / Q000235:genetics* / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C000715470:MAS1 protein, human; D000090063:Proto-Oncogene Mas; D000242:Cyclic AMP; D019009:Proto-Oncogene Proteins c-kit; D011493:Protein Kinase C
keywords: DASATINIB; HMC-1; IMATINIB; MAST CELLS; c-KIT RECEPTOR; cAMP/PKA/PATHWAY