@conference{
editor = "Kanazir, Selma, Savić, Danijela, Isaković, Aleksandra",
author = "Zogović, Nevena and Tovilović-Kovačević, Gordana and Misirkić Marjanović, Maja and Janjetović, Kristina and Vučićević, Ljubica and Trajković, Vladimir",
year = "2013",
abstract = "Introduction: Neural differentiation involves
intracellular signaling-controlled maturation of neural
progenitors into cells with fully developed neuronal
phenotype.
Aim: We explored the interplay between the
adenosine monophosphate-activated protein kinase
(AMPK), extracellular signal-regulated kinase (ERK) and
autophagy in phorbol myristate acetate (PMA)-induced
differentiation of SH-SY5Y human neuroblastoma cells.
Methods: The levels of neuronal marker expression
and acidification of autophagic compartments were
determined by flow cytometry of cells labeled with
appropriate antibodies and acridine orange, respectively.
The activation (phosphorylation) of AMPK and ERK, as
well as the levels of autophagic proteins beclin-1, Atg7,
microtubule-associated protein light chain 3 (LC3) and p62,
were assessed using immunoblot. RNA interference was
used for AMPK and LC3 knockdown.
Results: PMA initiated autophagic response in SH-
SY5Y cells simultaneously with activation of AMPK, a
major intracellular energy sensor, and ERK, a kinase
involved in cell proliferation and differentiation.
Pharmacological inhibition of AMPK or AMPK gene
silencing attenuated differentiation of SH-SY5Y cells, as
well as PMA-induced ERK activation and autophagy. A
selective pharmacological blockade of ERK prevented
PMA-induced neuronal differentiation and autophagy
induction, without affecting AMPK phosphorylation. On the
other hand, the inhibition of autophagy downstream of
AMPK/ERK activation, either by pharmacological agents or
LC3 knockdown, actually promoted the expression of
neuronal markers, thus indicating a role for autophagy in
suppression of PMA-induced differentiation of SH-SY5Y
cells.
Conclusion: PMA-induced differentiation of SH-
SY5Y cells depends on a complex interplay between
AMPK, ERK and autophagy, where AMPK and ERK
promote differentiation independently of autophagy, while
simultaneously inhibiting differentiation process through
autophagy-dependent mechanisms., Uvod: Diferencijacija neurona je proces
kontrolisan od strane nekoliko unutarćelijskih signala. U
toku diferencijacije dolazi do potpunog sazrevanja
progenitorskih ćelija u neurone.
Cilj: Cilj ovog rada je bio da se ispita uloga protein
kinaze regulisane adenozin monofosfatom (AMPK), kinaze
regulisane vanćelijskim signalima (ERK) i autofagije tokom
diferencijacije humanih neuroblastoma SH-SY5Y izazvane
forbol miristat acetatom (PMA).
Metode: Ekspresija neuronskih markera i
autofagija su praćene pomoću protočnog citofluorimetra
nakon bojenja ćelija sa odgovarajućim antitelima, odnosno
akridin oranžom. Aktivnosti AMPK, ERK, beklina, Atg7,
proteina lakog lanca 3 povezanog sa mikrotubulima (LC3),
p62, p70S6K i aktina su odreñene pomoću imunoblota.
Utišavanje AMPK i LC3 gena je izvedeno pomoću malih
interferirajućih RNK.
Rezultati: Istovremeno sa započinjanjem
autofagije, PMA je aktivirao AMPK protein, glavni
energetski senzor u ćeliji, i ERK kinazu, zaduženu za
regulaciju diferencijacije i proliferacije. Farmakološka
inhibicija AMPK i utišavanje gena za AMPK usporili su
diferencijaciju SH-SY5Y ćelija, i inhibirali su aktivaciju
ERK kinaze i autofagiju izazvane pomoću PMA. Blokiranje
aktivnosti ERK proteina pomoću specifičnog inhibitora
značajno je usporilo diferencijaciju SH-SY5Y ćelija i
autofagiju izazvane posredstvom PMA, bez uticaja na
aktivnost AMPK. Sa druge strane, inhibicija autofagije
nishodno od AMPK/ERK (farmakološka ili utišavanjem
gena za LC3) je povećala ekspresiju neuronskih markera,
što ukazuje na činjenicu da autofagija izazvana pomoću
PMA najverovatnije suprimira diferencijaciju SH-SY5Y
ćelija.
Zaključak: Diferencijaciju SH-SY5Y ćelija
izazvanu posredstvom PMA pospešuju AMPK i ERK
proteini nezavisno od autofagnog procesa. Istovremeno oba
molekula inhibiraju diferencijaciju preko mehanizama
zavisnih od autofagije.",
publisher = "Belgrade : Serbian Neuroscience Society",
journal = "6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia",
title = "AMPK and ERK regulate phorbol myristate acetate induced differentiation of SH-SY5Y neuroblastoma cells trough autophagy-dependent and –independent mehanisms, Protein kinaza regulisana adenozin monofosfatom i kinaza regulisana vanćelijskim signalima regulišu diferencijaciju SH-SY5Y ćelija neuroblastoma izazvanu forbol miristat acetatom preko mehanizama zavisnih i nezavisnih od autofagije",
pages = "58-58",
url = "https://hdl.handle.net/21.15107/rcub_ibiss_6354"
}
