ABSTRACTS OF THE 21st ANNUAL MEETING OF THE ITALIAN ...

Abstracts of the 21st Annual Meeting of the Italian Society of Uro-Oncology (SIUrO), 22-24 June, 2011, Naples, Italy
(Ypsilon Biotechnology, Napoli, Italy). Cells starved in red
phenol-free DMEM and 1% charcoal treated FBS for five
days were treated with 20 mM E 2 or 10 –6 or 10 –8 SOM230 or
20 mM E 2+ SOM230 (10 –8 or +10 –6 ) for 48 h. Cells were
differently harvested for semi-quantitative RT-PCR, Western
blot or flow cytometry (FACS) analyses. Results: E 2 induced
an up-regulation of SSR 1, 2 and 5 mRNA and proteins. E 2
and SOM 10 –6 alone induced apoptosis and slightly reduced
proliferation. The combined treatment of 20 mM E 2 +
SOM230 (10 –8 or 10 –6 ) induced a stronger rate of apoptosis
and a greater decrease of proliferation compared to either
alone. The synergistic action of SOM230 and E 2 induced a
reduction in S-phase proliferation with an arrest in G 0/G 1
phase. Caspase-dependent apoptosis was induced by
SOM230. while a reduction of BCL-2 levels was induced
after the addition of E 2, which amplified SOM230 effects at
lower doses. Conclusion: E 2 increases the inhibitory effects
of SOM230 in a prostate cell model expressing ER alpha and
beta, acting directly on cell growth and cell death control and
up-regulating SSRs.
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68
PROKINETICIN 1 STIMULATES THE
MIGRATION AND PROLIFERATION OF
PROSTATE EPITHELIAL CELLS IN VITRO
Giuseppe Bellastella1 , Valentina Rossi1 , Paolo Chieffi2 ,
Antonio Alberto Sinisi1 , Domenico Prezioso3 ,
Fabrizio Iacono3 , Ester Illiano3 ,
Carmine Cicalese3 and Raffaele Galasso3 1Endocrinologia, Dipartimento di Scienze Cardiotoraciche
and 2Dipartimento di Medicina Sperimentale, Seconda
Università di Napoli, Napoli, Italy;
3Clinica Urologia, Universitá Federico II Napoli, Napoli,
Italy
Background and Aim: Increased PROK1 expression has been
found in prostate hyperplasia and cancer, suggesting a role in
prostate proliferative diseases. The aim of this study was to
elucidate the role of PROK1 on prostate cell (PC) function and
growth. We evaluated the effects of PROK1 on epithelial PC
migration and proliferation using the androgen-dependent
epithelial PC line EPN, and a stabilized PC line derived from
prostate cancer (CPEC). Materials and Methods: Semiconfluent
starved cultures were treated with recombinant
PROK1 (5 nM), alone or associated with antiPROK1
monoclonal antibody or solvent. Cells were harvested 48 h after
the treatment and stained with propidium iodide for flow
cytometry of the cell cycle distribution by FACSCalibur or
recovered for protein extraction for Western blot analysis, or for
mRNA extraction for semi-quantitative RT-PCR. Cells grown
on slides were also treated and harvested after 46 h for TUNEL
assay. A wound assay was performed for the evaluation of cell
motility after overnight incubation. For ERK phosphorylation
assay, cell cultures were recovered after 5, 10. 20 and 60 min
following treatment. Results: An increase of the cell number in
S phase, with a decrease of cell counts in pre-G 1 and G 0/G 1 and
a significant reduction of the percentage of fragmented nuclei
was found after PROK1 treatment (p