Laredo J, Demur C, Muller C, Saivin S, Cassar G, Bousquet C, Dastugue N, Jaffrézou J P, Colombies P, Laurent G
Laboratoire de Pharmacologie et de Toxicologie Fondamentales, CNRS, Toulouse, France.
Leukemia. 1993 Jun;7(6):813-20.
In this study, we compared the impact of two protein kinase (PK) inhibitors, H-7 and staurosporine, on the normal myeloid progenitors (CFU-GM) and acute myeloid leukemia progenitors (AML-CFU) proliferation measured by in vitro clonogenic assay. H-7 and staurosporine displayed a biphasic dose-effect on both CFU-GM and AML-CFU recovery. At the lowest concentration range (0.1 microM to 20 microM for H-7 and 0.1 nM to 1 nM for staurosporine), we observed growth stimulation whereas higher concentrations induced dose-dependent growth inhibition. Moreover, AML-CFU proved to be significantly more sensitive to the inhibitory effect of both H-7 and staurosporine than CFU-GM (3.16- and 2.12-fold, respectively). These results were further confirmed with comparable murine cell line models (FDC-P1, a hematopoietic cell line generated from normal bone marrow and WEHI, a myelomonocytic leukemia cell line). Furthermore, we report that both H-7 and staurosporine present similar inhibitory effects on proliferation (PE1) as on self-renewal (PEs) of AML-CFU. In an attempt to understand more fully the mechanism of action of H-7 and staurosporine, we investigated their impact (when used at their D50) on the human myelogenous leukemia cell line, K562. H-7 and staurosporine induced a transient decrease of cell growth, between 0 and 24 hours, and produced a transient blockade of K562 cells in the S-phase, either 24 or 48 hours after the addition of staurosporine and H-7, respectively.
在本研究中，我们比较了两种蛋白激酶（PK）抑制剂H-7和星形孢菌素对通过体外克隆形成试验测量的正常髓系祖细胞（CFU-GM）和急性髓系白血病祖细胞（AML-CFU）增殖的影响。H-7和星形孢菌素对CFU-GM和AML-CFU的恢复均呈现双相剂量效应。在最低浓度范围（H-7为0.1微摩尔至20微摩尔，星形孢菌素为0.1纳摩尔至1纳摩尔），我们观察到生长刺激，而较高浓度则诱导剂量依赖性生长抑制。此外，事实证明，AML-CFU对H-7和星形孢菌素的抑制作用比CFU-GM敏感得多（分别为3.16倍和2.12倍）。使用可比的小鼠细胞系模型（FDC-P1，一种源自正常骨髓的造血细胞系和WEHI，一种髓单核细胞白血病细胞系）进一步证实了这些结果。此外，我们报告H-7和星形孢菌素对AML-CFU的增殖（PE1）和自我更新（PEs）具有相似的抑制作用。为了更全面地了解H-7和星形孢菌素的作用机制，我们研究了它们（在其半数效应浓度时使用）对人髓性白血病细胞系K562的影响。H-7和星形孢菌素分别在加入后24小时和48小时诱导细胞生长在0至24小时内短暂下降，并使K562细胞在S期短暂停滞。
