Makishima M, Honma Y, Hozumi M, Nagata N, Motoyoshi K
Department of Chemotherapy, Saitama Cancer Center Research Institute, Japan.
Biochim Biophys Acta. 1993 Apr 16;1176(3):245-9. doi: 10.1016/0167-4889(93)90051-p.
Inhibitors of protein kinase activities are useful for the study of intracellular signal transduction and some of these inhibitors are reported to induce differentiation of human leukemia cells. We examined effects of granulocyte-macrophage colony-stimulating factor (GM-CSF) in combination with several kinase inhibitors on differentiation of human leukemia U937 cells. Nitroblue tetrazolium (NBT)-reducing activity, a typical marker of myelomonocytic differentiation, of U937 cells was induced by genistein and GM-CSF enhanced this activity. GM-CSF also induced the NBT-reducing activity of the cells in combination with 2,5-dihydroxycinnamic acid methyl ester, psi-tectorigenin and staurosporine, although each of them did not induce the activity. Inhibitors of myosin light chain kinase, 1-(5-chloronaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (ML-9) and 1-(5-iodonaphthalene-1-sulfonyl)-1H-hexahydro-1,4-diazepine hydrochloride (ML-7), induced in U937 cells NBT-reduction, and lysozyme and alpha-naphthyl acetate esterase activities. GM-CSF inhibited this differentiation and counteracted the anti-proliferation effect of the kinase inhibitors. These results suggest that some protein kinases are involved in differentiation of U937 cells and the kinases inhibited by ML-9 and ML-7 are associated with signal transduction of GM-CSF.
蛋白激酶活性抑制剂对于研究细胞内信号转导很有用，并且据报道其中一些抑制剂可诱导人白血病细胞分化。我们研究了粒细胞-巨噬细胞集落刺激因子（GM-CSF）与几种激酶抑制剂联合使用对人白血病U937细胞分化的影响。染料木黄酮可诱导U937细胞的硝基蓝四氮唑（NBT）还原活性，这是骨髓单核细胞分化的典型标志物，GM-CSF可增强这种活性。GM-CSF与2,5-二羟基肉桂酸甲酯、次甘草素和星形孢菌素联合使用时也可诱导细胞的NBT还原活性，尽管它们各自单独使用时不能诱导该活性。肌球蛋白轻链激酶抑制剂1-(5-氯萘-1-磺酰基)-1H-六氢-1,4-二氮杂䓬盐酸盐（ML-9）和1-(5-碘萘-1-磺酰基)-1H-六氢-1,4-二氮杂䓬盐酸盐（ML-7）可诱导U937细胞的NBT还原以及溶菌酶和α-萘乙酸酯酶活性。GM-CSF抑制这种分化并抵消激酶抑制剂的抗增殖作用。这些结果表明，一些蛋白激酶参与U937细胞的分化，并且被ML-9和ML-7抑制的激酶与GM-CSF的信号转导有关。
