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,G.?Ramponi author-information" > author-information__contact u-icon-before" > mailto:ramponi@scibio.unifi.it" title=" ramponi@scibio.unifi.it" itemprop=" email" data-track=" click" data-track-action=" Email author" data-track-label=" " >Email author |
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Affiliation: | (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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