Histamine actions in the normal human fibroblast cell line CCD-1059Sk

PORRETTI J, MOHAMAD N, BADENAS E, ESNAOLA M, RIVERA E, MARTÍN G, CRICCO G

Belfast

Congreso; XLI Annual Meeting of the European Histamine Research Society (EHRS); 2012

European Histamine Research Society (EHRS)

We have previously demonstrated a high expression of histidine decarboxilase (HDC), the enzyme involved in histamine (HA) synthesis and therefore a high HA content in the human pancreatic carcinoma cell line BxPC3. In vivo, HA produced a significant desmoplasia in nude mice bearing BxPC3 xenografts and also neoplastic cells colonizing lung parenchyma were observed. In vitro, conditioned medium of BxPC3 cells treated with (HA 1-10 uM) increased the number of CCD-1059Sk cells in S phase of cell cycle. The aim of this work was to evaluate HA action on cell proliferation, gelatinases expression and activity and cell motility in the normal human fibroblast cell line CCD-1059Sk. A direct action of HA (1-10 uM) on fibroblast proliferation was observed employing bromodeoxyuridine (BrdU) incorporation technique. After 8h of HA treatment there was a significant rise in fibroblast cells number in S phase (p<0.05 vs non-treated cells). By RT-PCR we determined the mRNA expression of H1, H2, H3 and H4 histamine receptors, the matrix metalloproteinase 2 (MMP2) and the tissue inhibitor of MMP (TIMP1) in this cell line. Immunostaining studies showed that CCD-1059Sk fibroblasts express cytoplasmatic MMP2 protein, though no difference in MMP2 expression was observed between HA treated and non-treated cells. Gelatinolytic activity evaluated by zymography was significantly increased by 10 uM HA and reproduced by the H1 histamine receptor agonist. 10 uM HA enhanced cell migration (p<0.05 vs non treated cells) assessed using transwell units. In summary, we may conclude that exogenous HA exerts a direct action on the normal human fibroblasts CCD-1059Sk proliferation, gelatinolytic activity and motility, opening the perspective for future studies on the role of histamine in fibroblasts activation during the epithelial mesenchymal transition related to cancer progression.