The protein kinase C inhibitor H7 blocks phosphorylation of stathmin during TPA-induced growth inhibition of human pre-B leukemia REH6 cells.
The human pre-B acute lymphoblastic leukemia cell line REH6 was used to analyze the regulation of a ubiquitous intracellular phosphoprotein stathmin (Mr 19,000, pl = 5.6-6.2). We demonstrated by 32P-labeling that the short (1 h) treatment of the REH6 cells with the tumor promoting phorbol ester, 12-O-tetradecanoylphorbol-13-acetate (TPA), resulted in a rapid phosphorylation of at least three (P1, P2 and P3) stathmin isoforms without an alteration of stathmin isoform expression. Furthermore, Western blot analysis with specific antiserum showed that the prolonged period (48 h) of TPA treatment partially reduced protein levels particularly of two (N2 and P2) stathmin isoforms. The potent and relatively specific protein kinase C (PKC) inhibitor, 1,(5-isoquinolinesulphonyl)2methylpiperasine dihydrochloride (H7), partially inhibited these TPA effects, whereas the specific calmodulin inhibitor R24571 (calmidazolium) had no effect upon these events. Our findings suggest that stathmin phosphorylation in REH6 cells could be in part mediated by PKC activation.
Duraj
J
J
Department of Molecular Immunology, Slovak Academy of Sciences, Bratislava.
Kovacikova
M
M
Sedlak
J
J
Koppel
J
J
Sobel
A
A
Chorvath
B
B
D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D002051:Burkitt Lymphoma / Q000378:metabolism / Q000473:pathology*; D002147:Calmodulin / Q000037:antagonists & inhibitors; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology; D007546:Isoquinolines / Q000494:pharmacology*; D008868:Microtubule Proteins; D010750:Phosphoproteins / Q000378:metabolism*; D010766:Phosphorylation; D010879:Piperazines / Q000494:pharmacology*; D011289:Preleukemia / Q000378:metabolism / Q000473:pathology*; D011493:Protein Kinase C / Q000037:antagonists & inhibitors*; D050777:Stathmin; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology*; D014407:Tumor Cells, Cultured
D002147:Calmodulin; D007093:Imidazoles; D007546:Isoquinolines; D008868:Microtubule Proteins; D010750:Phosphoproteins; D010879:Piperazines; C493995:STMN1 protein, human; D050777:Stathmin; C031938:calmidazolium; D019307:1-(5-Isoquinolinesulfonyl)-2-Methylpiperazine; D011493:Protein Kinase C; D013755:Tetradecanoylphorbol Acetate