Histamine modulates BxPC3 cell line growth and the interaction with the microenvironment

CRICCO GRACIELA; MOHAMAD NORA; PORRETTI JULIANA; RIVERA ELENA; MARTIN GABRIELA

Sochi

Congreso; XL European Histamine Research Society Meeting; 2011

European Histamine Research Society

We have previously demonstrated that histamine (HA) regulates processes related to cell proliferation, differentiation and invasive ability via the four membrane receptors in various tumor cell lines. The aim of this work was to assess the role HA in a human pancreatic adenocarcinoma cell line BxPC3. We determined that HA modulates cell proliferation in a concentration dependent way using a clonogenic assay. A rise was observed at low doses of HA (0.5 uM-1 uM) while a decrease was found over 20 uM). The stimulatory effect was evoked by the H3 agonist (R)-á-Methylhistamine and the H2 agonist Amthamine; both responses were blocked by the specific antagonists Ranitidine and JNJ5207852. Bromodeoxiuridin (BrdU) incorporation evaluated by immunostaining and cyclin D and E expression by immunoblot confirmed the increase in the number of cells in S phase of the cell cycle when cultures were treated with low doses of HA or Amthamine or (R)-á-Methylhistamine. Conversely the H1 agonist 2-(3-trifluormethylphenyl)histamine and H4 agonists VUF 8430 and Clobenpropit inhibited cell proliferation by increasing apoptosis. Augmented matrix metalloproteinases secretion and cell migration are critical for tumor invasion. We found that HA enhances dose dependently (0.5-25 uM) MMP9 activity (assessed by zymography) and cell migration (evaluated by transwell systems). Subcutaneous inoculation of BxPC3 cells in nude mice led to the development of differentiated tumors with cystic features. Tumors xenografted in HA treated animals showed a higher growth rate and an important stromal desmoplastic reaction than tumors in control animals. In vitro studies determined that conditioned media from HA treated BxPC3 cells, increased proliferation of a normal fibroblast cell line, assayed by BrdU incorporation and PCNA expression.Pancreatic cancer is a disease with a high death rate; the identification of drugs that can both regulate tumor cell survival and metastatic ability will help to delineate more effective strategies for therapeutic intervention.