Skip to Content
Merck
  • p62/SQSTM1 upregulation constitutes a survival mechanism that occurs during granulocytic differentiation of acute myeloid leukemia cells.

p62/SQSTM1 upregulation constitutes a survival mechanism that occurs during granulocytic differentiation of acute myeloid leukemia cells.

Cell death and differentiation (2014-07-19)
A Trocoli, P Bensadoun, E Richard, G Labrunie, F Merhi, A M Schläfli, D Brigger, S Souquere, G Pierron, J-M Pasquet, P Soubeyran, J Reiffers, E Ségal-Bendirdjian, M P Tschan, M Djavaheri-Mergny
ABSTRACT

The p62/SQSTM1 adapter protein has an important role in the regulation of several key signaling pathways and helps transport ubiquitinated proteins to the autophagosomes and proteasome for degradation. Here, we investigate the regulation and roles of p62/SQSTM1 during acute myeloid leukemia (AML) cell maturation into granulocytes. Levels of p62/SQSTM1 mRNA and protein were both significantly increased during all-trans retinoic acid (ATRA)-induced differentiation of AML cells through a mechanism that depends on NF-κB activation. We show that this response constitutes a survival mechanism that prolongs the life span of mature AML cells and mitigates the effects of accumulation of aggregated proteins that occurs during granulocytic differentiation. Interestingly, ATRA-induced p62/SQSTM1 upregulation was impaired in maturation-resistant AML cells but was reactivated when differentiation was restored in these cells. Primary blast cells of AML patients and CD34(+) progenitors exhibited significantly lower p62/SQSTM1 mRNA levels than did mature granulocytes from healthy donors. Our results demonstrate that p62/SQSTM1 expression is upregulated in mature compared with immature myeloid cells and reveal a pro-survival function of the NF-κB/SQSTM1 signaling axis during granulocytic differentiation of AML cells. These findings may help our understanding of neutrophil/granulocyte development and will guide the development of novel therapeutic strategies for refractory and relapsed AML patients with previous exposure to ATRA.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Anti-LC3B antibody produced in rabbit, ~1 mg/mL, affinity isolated antibody, buffered aqueous solution
Sigma-Aldrich
8-(4-Chlorophenylthio)adenosine 3′,5′-cyclic monophosphate sodium salt, ≥97.0% (HPLC), powder
Sigma-Aldrich
Retinoic acid, ≥98% (HPLC), powder
Sigma-Aldrich
Puromycin dihydrochloride from Streptomyces alboniger, powder, BioReagent, suitable for cell culture
Sigma-Aldrich
Propidium iodide, ≥94% (HPLC)
Sigma-Aldrich
Propidium iodide, ≥94.0% (HPLC)
USP
Tretinoin, United States Pharmacopeia (USP) Reference Standard
Supelco
Tretinoin, Pharmaceutical Secondary Standard; Certified Reference Material
Tretinoin, European Pharmacopoeia (EP) Reference Standard
Sigma-Aldrich
MG-132, Ready Made Solution, ≥90% (HPLC)
Sigma-Aldrich
Triton X-100, laboratory grade
Sigma-Aldrich
Tetramethylrhodamine methyl ester perchlorate, ≥95%
Sigma-Aldrich
Z-Leu-Leu-Leu-al, ≥90% (HPLC)
Sigma-Aldrich
Propidium iodide solution
Sigma-Aldrich
Anti-GTF2H1 antibody produced in rabbit, IgG fraction of antiserum