Konference: 2007 49th ASH Annual Meeting - účast ČR
Kategorie:
Maligní lymfomy a leukémie
Téma: Publikace ve sborníku
Číslo abstraktu: 4155
Autoři: Doc.MUDr. Jan Živný, Ph.D.; prof. MUDr. Pavel Klener, DrSc.; Mgr. Ondřej Toman, Ph.D.; Ing. Petr Halada, Ph.D.; Robert Ivanek; Mgr. Tereza Šimonová; RNDr. Ladislav Anděra, CSc.; Prof. MUDr. Emanuel Nečas, DrSc.; RNDr. Jiří Petrák, Ph.D.
Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL or
Apo2L) is a proapoptotic cytokine implicated in cancer cell
surveillance. TRAIL induces apoptosis of target cells by the
extrinsic, receptor-mediated, apoptotic pathway. A potential of
TRAIL as cancer-specific therapeutic agent has been proposed,
either as a single agent or in combination with chemotherapy
agents. Development of TRAIL-resistant clones in the originally
TRAIL-sensitive tumor cell population may be a critical
complication of TRAIL based cancer therapy. Prolonged exposure of
TRAIL-sensitive leukemia cell line, wild-type (WT) HL60 cells, to
recombinant soluble TRAIL resulted in the establishment of
resistant subclones. To get deeper insight into the molecular
mechanism of acquired TRAIL resistance we compared expression
profiles of TRAIL-sensitive HL-60 cells and TRAIL-resistant HL-60
subclones using microarray mRNA gene expression chips and 2D-PAGE
followed by MS and MS/MS analysis. Using cluster analysis of
microarray mRNA gene expression profiles of 6 individual HL60
resistant subclones we identified two molecular signatures of TRAIL
resistance. When compared to WT HL60 TRAIL-sensitive mRNA
expression the group of HL60 subclones designed as phenotype 1 (P1,
n=3) had 1122 significantly upregulated and 985 significantly
downregulated mRNAs and subclones designed as phenotype 2 (P2, n=3)
had 1583 significantly upregulated and 1798 significantly
downregulated mRNAs (p<0.05). The subclones of P1 and P2
phenotype shared 533 upregulated and 422 downregulated genes
compared to HL60 WT cells. To identify differentially expressed
proteins we performed 2D-PAGE of two representative subclones of P1
and P2 phenotypes (in five replicates) and compared them with HL60
WT cells. We detected and identified differentially expressed
proteins involved in various aspects of cellular metabolism. Except
for downregulation of chromobox protein homolog 5, a protein
involved in the regulation of gene expression, and downregulation
of Annexin A6 and protein disulfide-isomerase A3 precursor the
TRAIL-resistant P1 and P2 subclones showed different protein
expression profile. In P1 TRAIL-resistant cells we identified
downregulation of proteins involved in energy metabolism, including
pyruvate kinase, enolase, cytochrome c-reductase, NADH
dehydrogenase and downregulation of proteins involved in proteasome
activity, such as proteasome activator complex subunits 1 and 2. In
P2 TRAIL-resistant cells we detected downregulation of important
regulatory proteins, such as DNA replication licensing factor MCM7,
proliferation-associated protein 2G4, replication protein A 32 kDa
subunit. In both, P1 and P2, subclones we identified significant
changes in cytoskeletal rearrangement proteins. In addition,
several chaperones were differentially expressed, namely
protein-disulfide isomerase, GrpE, HSC71 and HSP27, 78 kDa
glucose-regulated protein precursor. Our data show two HL60 cell
line-derived TRAIL resistant subclones, P1 and P2, have specific
molecular signatures suggesting distinct mechanisms of TRAIL
resistance. These distinct pathways for development of resistance
to TRAIL-induced apoptosis are relevant for design of more
effective strategies for leukemia therapy. Supported by MZCR
023736, MSM LC06044 and 0021620806, and IGA MZ NR8317 and NR8930.
Abstract #4155 appears in Blood, Volume 110, issue 11, November 16,
2007
Keywords: Microarray Analysis|Proteomics|Drug Resistance
Disclosure: No relevant conflicts of interest to declare.
Session Info: Publication Only
Datum přednesení příspěvku: 8. 12. 2007