Hu Z B, Ma W, Uphoff C C, Quentmeier H, Drexler H G
DSM-German Collection of Microorganisms and Cell Cultures, Department of Human and Animal Cell Cultures, Braunschweig.
Blood. 1994 Apr 15;83(8):2133-44.
A panel of 164 continuous human leukemia-lymphoma cell lines was analyzed for expression of c-kit using Northern blotting and reverse transcriptase-polymerase chain reaction (RT-PCR). The c-kit transcripts were detectable in cell lines assigned to the myeloid (in 7 of 29 by Northern blotting and in 4 of 8 by RT-PCR), monocytic (in 1 of 24 by Northern blotting and in 3 of 6 by RT-PCR), erythroid (in 6 of 8 by Northern blotting and in 5 of 5 by RT-PCR), and megakaryoblastic (in 10 of 10 by Northern blotting) lineages, c-kit expression was not seen by Northern blotting or RT-PCR analysis in any of the 93 lymphoid leukemia, myeloma, or lymphoma cell lines. Treatment of four megakaryoblastic cell lines with protein kinase C activators (phorbol ester 12-O-tetradecanoylphorbol 13-acetate and Bryostatin 1) led to terminal differentiation as assessed by morphologic alterations, changes in the surface marker profile, and growth arrest. These effects were associated with enhanced c-kit mRNA expression. Exposure to all-trans retinoic acid down-regulated c-kit mRNA levels, while simultaneously causing morphologic alterations in all four cell lines. Stimulation with growth factors (interleukin-3, granulocyte macrophage-colony stimulating factor, and insulin-like growth factors I and II), used to assess any role of c-kit in proliferative processes, did not lead to significant upregulation or downregulation of c-kit expression. The finding of constitutive and high expression of c-kit mRNA in all megakaryoblastic leukemia cell lines and its modulation by various reagents might further contribute to the understanding of megakaryopoietic proliferation, differentiation, and leukemogenesis.
使用Northern印迹法和逆转录聚合酶链反应(RT-PCR)分析了164种连续的人类白血病-淋巴瘤细胞系的c-kit表达情况。在归类于髓系(Northern印迹法检测29种中的7种,RT-PCR检测8种中的4种)、单核细胞系(Northern印迹法检测24种中的1种,RT-PCR检测6种中的3种)、红系(Northern印迹法检测8种中的6种,RT-PCR检测5种中的5种)以及巨核母细胞系(Northern印迹法检测10种中的10种)的细胞系中可检测到c-kit转录本。在93种淋巴样白血病、骨髓瘤或淋巴瘤细胞系中,通过Northern印迹法或RT-PCR分析均未观察到c-kit表达。用蛋白激酶C激活剂(佛波酯12-O-十四烷酰佛波醇13-乙酸酯和苔藓抑素1)处理四种巨核母细胞系,通过形态学改变、表面标志物谱变化和生长停滞评估发现其导致了终末分化。这些效应与c-kit mRNA表达增强相关。暴露于全反式维甲酸会下调c-kit mRNA水平,同时在所有四种细胞系中引起形态学改变。用生长因子(白细胞介素-3、粒细胞巨噬细胞集落刺激因子以及胰岛素样生长因子I和II)刺激以评估c-kit在增殖过程中的任何作用,并未导致c-kit表达的显著上调或下调。在所有巨核母细胞白血病细胞系中发现c-kit mRNA的组成性高表达及其受各种试剂的调节,可能会进一步有助于理解巨核细胞生成、分化和白血病发生。