RRC ID 30713
Author Nagao T, Kurosu T, Umezawa Y, Nogami A, Oshikawa G, Tohda S, Yamamoto M, Miura O.
Title Proliferation and survival signaling from both Jak2-V617F and Lyn involving GSK3 and mTOR/p70S6K/4EBP1 in PVTL-1 cell line newly established from acute myeloid leukemia transformed from polycythemia vera.
Journal PLoS ONE
Abstract The gain of function mutation JAK2-V617F is very frequently found in myeloproliferative neoplasms (MPNs) and is strongly implicated in pathogenesis of these and other hematological malignancies. Here we report establishment of a new leukemia cell line, PVTL-1, homozygous for JAK2-V617F from a 73-year-old female patient with acute myeloid leukemia (AML) transformed from MPN. PVTL-1 is positive for CD7, CD13, CD33, CD34, CD117, HLA-DR, and MPO, and has complex karyotypic abnormalities, 44,XX,-5q,-7,-8,add(11)(p11.2),add(11)(q23),-16,+21,-22,+mar1. Sequence analysis of JAK2 revealed only the mutated allele coding for Jak2-V617F. Proliferation of PVTL-1 was inhibited and apoptosis was induced by the pan-Jak inhibitor Jak inhibitor-1 (JakI-1) or dasatinib, which inhibits the Src family kinases as well as BCR/ABL. Consistently, the Src family kinase Lyn was constitutively activated with phosphorylation of Y396 in the activation loop, which was inhibited by dasatinib but not by JakI-1. Further analyses with JakI-1 and dasatinib indicated that Jak2-V617F phosphorylated STAT5 and SHP2 while Lyn phosphorylated SHP1, SHP2, Gab-2, c-Cbl, and CrkL to induce the SHP2/Gab2 and c-Cbl/CrkL complex formation. In addition, JakI-1 and dasatinib inactivated the mTOR/p70S6K/4EBP1 pathway and reduced the inhibitory phosphorylation of GSK3 in PVTL-1 cells, which correlated with their effects on proliferation and survival of these cells. Furthermore, inhibition of GSK3 by its inhibitor SB216763 mitigated apoptosis induced by dasatinib but not by JakI-1. Together, these data suggest that apoptosis may be suppressed in PVTL-1 cells through inactivation of GSK3 by Lyn as well as Jak2-V617F and additionally through activation of STAT5 by Jak2-V617F. It is also speculated that activation of the mTOR/p70S6K/4EBP1 pathway may mediate proliferation signaling from Jak2-V617F and Lyn. PVTL-1 cells may provide a valuable model system to elucidate the molecular mechanisms involved in evolution of Jak2-V617F-expressing MPN to AML and to develop novel therapies against this intractable condition.
Volume 9(1)
Pages e84746
Published 2014
DOI 10.1371/journal.pone.0084746
PII PONE-D-13-34652
PMID 24404189
PMC PMC3880321
MeSH Adaptor Proteins, Signal Transducing / metabolism* Apoptosis / drug effects Bone Marrow / metabolism Bone Marrow / pathology Cell Line, Tumor Cell Proliferation / drug effects Cell Survival / drug effects Cell Survival / genetics Cell Transformation, Neoplastic / genetics Cell Transformation, Neoplastic / metabolism DNA Mutational Analysis Dasatinib Female Glycogen Synthase Kinase 3 / metabolism* Humans Janus Kinase 2 / genetics Janus Kinase 2 / metabolism* Karyotype Leukemia, Myeloid, Acute / diagnosis Leukemia, Myeloid, Acute / genetics Leukemia, Myeloid, Acute / metabolism* Middle Aged Mutation Phosphoproteins / metabolism* Phosphorylation / drug effects Polycythemia Vera / diagnosis Polycythemia Vera / genetics Polycythemia Vera / metabolism* Protein Kinase Inhibitors / pharmacology Pyrimidines / pharmacology Ribosomal Protein S6 Kinases, 70-kDa / metabolism* Signal Transduction / drug effects Thiazoles / pharmacology src-Family Kinases / genetics src-Family Kinases / metabolism*
IF 2.766
Times Cited 6
WOS Category HEMATOLOGY
Resource
DNA material pRx nZ ires Neo (RDB01699).