Expression of protein kinase c (pkc) isoforms and the effect of selective pkc-s inhibition by rottlerin on the induction of apoptosis in acute promyelocytic leukemia

Hesham Amin, Melek Ergin, Mitchell F. Denning, Shahnaz Saeed, Serhan Alkan

Research output: Contribution to journalArticle

Abstract

Acute promyelocytic leukemia (APL; FAB-M3) is a distinct form of acute leukemia, characterized by specific r(15;17) chromosomal translocation and distinct morphologic features. In a recent report, we demonstrated that inhibition of protein kinase C (PKC) induces significant apoptosis in APL cells from patients and cell-lines (fir. J. Haematol. 2000; 110:552-562). The PKC family consists of at least 11 different isoforms, which have been characterized and divided into three major groups based on structural and functional properties. These groups are the conventional or Ca2 -dependent (cPKC): a, , and y, the novel or Ca2Mndependent (nPKC): 8, e, r\, 8, and n; and the atypical (aPKC): Ç, X, and iota. Using western blot analysis, we studied the expression pattern of the different PKC isoforms in 5 APL patients with r(15;17) translocation, and also in two APL cell-lines, NB4 and its all-rrons-retinoic acid (ATRA)-resistant counterpart NB4.306. The expression of the PKC isoforms in primary APL cells and cell-lines is illustrated in the table: α β γ δ ε λ θ NB4 + + + + + - + + NB4.306 + + + - + + Patient #1 - - - + - - + Patient #2 + + + + - - + Patient #3 + + + + - + - Patient #4 + + + + + - + Patient #5 + + - + - + + We also investigated the apoptotic effects of PKC-5 selective inhibition by rottlerin (10 mM at 72 h). Inhibition of PKC-5 by rottlerin induced greater than 50% apoptosis in primary APL cells and NB4 and NB4.306 cell-lines, as documented by flow cytometric analysis (annexin-7AAD) and morphologic changes characteristic of apoptosis. In conclusion, there is a variable expression of PKC isoforms in primary APL cells and celllines. The consistent expression of PKC-8 along with the finding of significant rottlerininduced apoptosis suggest that PKC-8 is an important isoform for survival of APL cells. These results also indicate that utilization of selective inhibitors of specific PKC isoforms could be a potential therapeutic modality in APL patients.

Original languageEnglish (US)
JournalBlood
Volume96
Issue number11 PART II
StatePublished - Dec 1 2000

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Acute Promyelocytic Leukemia
Protein Kinases
Protein Kinase C
Protein Isoforms
Apoptosis
Cell Line
rottlerin
Annexins
Genetic Translocation
Tretinoin
Cell Survival
Leukemia
Western Blotting

ASJC Scopus subject areas

  • Biochemistry
  • Immunology
  • Hematology
  • Cell Biology

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Expression of protein kinase c (pkc) isoforms and the effect of selective pkc-s inhibition by rottlerin on the induction of apoptosis in acute promyelocytic leukemia. / Amin, Hesham; Ergin, Melek; Denning, Mitchell F.; Saeed, Shahnaz; Alkan, Serhan.

In: Blood, Vol. 96, No. 11 PART II, 01.12.2000.

Research output: Contribution to journalArticle

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title = "Expression of protein kinase c (pkc) isoforms and the effect of selective pkc-s inhibition by rottlerin on the induction of apoptosis in acute promyelocytic leukemia",
abstract = "Acute promyelocytic leukemia (APL; FAB-M3) is a distinct form of acute leukemia, characterized by specific r(15;17) chromosomal translocation and distinct morphologic features. In a recent report, we demonstrated that inhibition of protein kinase C (PKC) induces significant apoptosis in APL cells from patients and cell-lines (fir. J. Haematol. 2000; 110:552-562). The PKC family consists of at least 11 different isoforms, which have been characterized and divided into three major groups based on structural and functional properties. These groups are the conventional or Ca2 -dependent (cPKC): a, , and y, the novel or Ca2Mndependent (nPKC): 8, e, r\, 8, and n; and the atypical (aPKC): {\cC}, X, and iota. Using western blot analysis, we studied the expression pattern of the different PKC isoforms in 5 APL patients with r(15;17) translocation, and also in two APL cell-lines, NB4 and its all-rrons-retinoic acid (ATRA)-resistant counterpart NB4.306. The expression of the PKC isoforms in primary APL cells and cell-lines is illustrated in the table: α β γ δ ε λ θ NB4 + + + + + - + + NB4.306 + + + - + + Patient #1 - - - + - - + Patient #2 + + + + - - + Patient #3 + + + + - + - Patient #4 + + + + + - + Patient #5 + + - + - + + We also investigated the apoptotic effects of PKC-5 selective inhibition by rottlerin (10 mM at 72 h). Inhibition of PKC-5 by rottlerin induced greater than 50{\%} apoptosis in primary APL cells and NB4 and NB4.306 cell-lines, as documented by flow cytometric analysis (annexin-7AAD) and morphologic changes characteristic of apoptosis. In conclusion, there is a variable expression of PKC isoforms in primary APL cells and celllines. The consistent expression of PKC-8 along with the finding of significant rottlerininduced apoptosis suggest that PKC-8 is an important isoform for survival of APL cells. These results also indicate that utilization of selective inhibitors of specific PKC isoforms could be a potential therapeutic modality in APL patients.",
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T1 - Expression of protein kinase c (pkc) isoforms and the effect of selective pkc-s inhibition by rottlerin on the induction of apoptosis in acute promyelocytic leukemia

AU - Amin, Hesham

AU - Ergin, Melek

AU - Denning, Mitchell F.

AU - Saeed, Shahnaz

AU - Alkan, Serhan

PY - 2000/12/1

Y1 - 2000/12/1

N2 - Acute promyelocytic leukemia (APL; FAB-M3) is a distinct form of acute leukemia, characterized by specific r(15;17) chromosomal translocation and distinct morphologic features. In a recent report, we demonstrated that inhibition of protein kinase C (PKC) induces significant apoptosis in APL cells from patients and cell-lines (fir. J. Haematol. 2000; 110:552-562). The PKC family consists of at least 11 different isoforms, which have been characterized and divided into three major groups based on structural and functional properties. These groups are the conventional or Ca2 -dependent (cPKC): a, , and y, the novel or Ca2Mndependent (nPKC): 8, e, r\, 8, and n; and the atypical (aPKC): Ç, X, and iota. Using western blot analysis, we studied the expression pattern of the different PKC isoforms in 5 APL patients with r(15;17) translocation, and also in two APL cell-lines, NB4 and its all-rrons-retinoic acid (ATRA)-resistant counterpart NB4.306. The expression of the PKC isoforms in primary APL cells and cell-lines is illustrated in the table: α β γ δ ε λ θ NB4 + + + + + - + + NB4.306 + + + - + + Patient #1 - - - + - - + Patient #2 + + + + - - + Patient #3 + + + + - + - Patient #4 + + + + + - + Patient #5 + + - + - + + We also investigated the apoptotic effects of PKC-5 selective inhibition by rottlerin (10 mM at 72 h). Inhibition of PKC-5 by rottlerin induced greater than 50% apoptosis in primary APL cells and NB4 and NB4.306 cell-lines, as documented by flow cytometric analysis (annexin-7AAD) and morphologic changes characteristic of apoptosis. In conclusion, there is a variable expression of PKC isoforms in primary APL cells and celllines. The consistent expression of PKC-8 along with the finding of significant rottlerininduced apoptosis suggest that PKC-8 is an important isoform for survival of APL cells. These results also indicate that utilization of selective inhibitors of specific PKC isoforms could be a potential therapeutic modality in APL patients.

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