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Survival role of protein kinase C (PKC) in chronic lymphocytic leukemia and determination of isoform expression pattern and genes altered by PKC inhibition
Author(s) -
Alkan Serhan,
Huang Qin,
Ergin Melek,
Denning Mitchell F.,
Nand Sucha,
Maududi Tazeen,
Paner Gladell P.,
Ozpuyan Fulya,
Izban Keith F.
Publication year - 2005
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/ajh.20352
Subject(s) - protein kinase c , gene isoform , chronic lymphocytic leukemia , microbiology and biotechnology , biology , western blot , gene expression , microarray analysis techniques , blot , downregulation and upregulation , cancer research , apoptosis , northern blot , signal transduction , gene , leukemia , immunology , genetics
Recent studies have suggested that protein kinase C (PKC) activation plays an important role in survival of chronic lymphocytic leukemia (CLL). In order to characterize the role of PKC in CLL, we investigated the expression pattern of PKC isoforms in CLL cells (7 cases) and evaluated the effect of PKC inhibition on the survival of CLL cells (20 cases). Expression of the classical PKC isoforms β and γ, the novel isoform δ and the atypical isoform ζ was seen in all analyzed patient samples by Western blot analysis. Expression of the PKC isoforms α, ε, and ι was variable. Following incubation with the PKC inhibitor, safingol, CLL cells underwent marked apoptosis in all cases. In order to characterize the molecular events associated with the apoptotic effect of PKC inhibition, gene expression patterns in CLL cells were evaluated by cDNA‐microarray analysis. Following safingol treatment, several genes showed marked downregulation and PKC‐related proteins demonstrated decreased hybridization signals. Among these proteins, CREB and Daxx were further studied by using Western blotting, nuclear binding assay and confocal immunofluorescent microscopy. These studies showed significant inhibition of these proteins, consistent with the results of microarray gene analysis. Overall, these findings suggest that PKC activation is important for CLL cell survival and that inhibitors of PKC may have a role in the treatment of patients with CLL. Am. J. Hematol. 79:97–106, 2005. © 2005 Wiley‐Liss, Inc.