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
2013
Authors:
Zogović, NevenaTovilović-Kovačević, Gordana
Misirkić Marjanović, Maja
Janjetović, Kristina
Vučićević, Ljubica
Trajković, Vladimir
Contributors
Kanazir, SelmaSavić, Danijela
Isaković, Aleksandra
Document Type:
Conference object (Published version)
,
© 2013 by the Serbian Neuroscience Society
Metadata
Show full item recordAbstract:
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.
Keywords:
neuronal differentiation; autophagy; AMPK; ERKFunding / projects:
- Modulation of intracellular energy balance-controlling signalling pathways in therapy of cancer and neuro-immuno-endocrine disorders (RS-MESTD-Integrated and Interdisciplinary Research (IIR or III)-41025)
- The role of autophagy in regulation of cancer cell death (RS-MESTD-Basic Research (BR or ON)-173053)
In:
- Kanazir S, Savić D, Isaković A, editors. 6th SNS Serbian Neuroscience Society Congress: book of abstracts; 2013 Nov 14-16; Belgrade, Serbia. Belgrade: Serbian Neuroscience Society; 2013. p. 58.