Acylphosphatase overexpression triggers SH-SY5Y differentiation towards neuronal phenotype |
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Authors: | C?Cecchi G?Liguri C?Fiorillo F?Bogani M?Gambassi E?Giannoni P?Cirri S?Baglioni Email author" target="_blank">G?RamponiEmail author |
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Institution: | (1) Department of Biochemical Sciences, University of Florence, Viale Morgagni 50, 50134 Florence, Italy |
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Abstract: | An acylphosphatase (AcPase) overexpression study was carried out on SH-SY5Y neuroblastoma cells, using a
green fluorescent fusion protein (AcP-GFP), with GFP acting as a reporter protein. The cellular proliferation rate
was significantly reduced by overexpression of AcPase by a factor of ten. In contrast, clones transfected with two
inactive AcPase mutants showed a growth rate comparable to control cells. This suggests that AcPase catalyzes the
proliferative down-regulation. AcPase-overexpressing clones showed a physiological mortality rate as assessed by an
MTT reduction test and by evaluation of necrotic markers. DNA fragmentation analysis and assays of caspase-3 and
poly (ADP-ribose) polymerase (PARP)-active fragments showed no evidence of any apoptotic pattern. AcPase
overexpression led to a marked increase in PARP activity as well as Bcl-2 content; these are commonly up-regulated
during differentiative processes in neuronal cells. In fact, the typical differentiation marker,
growth-associated-protein 43, was significantly up-regulated. Microscopic observations also showed a clear
increase in the differentiative phenotype in AcPase-overexpressing cells. Our results clearly show that AcPase
plays a primary causative role in neuronal differentiation.Received 3 May 2004; accepted 25 May 2004 |
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Keywords: | Acylphosphatase SH-SY5Y neuroblastoma cell differentiation apoptosis PMA |
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