Solomon D H, O'Driscoll K, Sosne G, Weinstein I B, Cayre Y E
Department of Physiology, College of Physicians and Surgeons, Columbia University, New York, New York 10032.
Cell Growth Differ. 1991 Apr;2(4):187-94.
The human promyelocytic leukemia cell line HL-60 differentiates in vitro when treated with various inducers. It has previously been shown that protein kinase C (PKC) isozymes are modulated during granulocytic differentiation of HL-60 cells induced by dimethyl sulfoxide or retinoic acid (M. Makowske, R. Ballester, Y. Cayre, and O.M. Rosen, J. Biol. Chem., 263: 3402-3410, 1988; K. Hashimoto, A. Kishimoto, H. Aihara, I. Yasuda, K. Mikawa, and Y. Nishizuka, FEBS Left., 263: 31-34, 1990). HL-60 responds to 1 alpha, 25-dihydroxyvitamin D3 (1,25-(OH)2D3) or to 12-O-tetradecanoylphorbol-13-acetate by giving rise to monocytic cells. In the present study, we demonstrate that treatment of HL-60 cells with 1,25-(OH)2D3 causes dramatic increases in PKC-alpha and PKC-beta protein levels detected by immunoblotting with PKC isoform-specific antibodies and in Ca(2+)- and phospholipid-dependent protein kinase activity. We also observed a transient increase in the steady-state levels of PKC-alpha and PKC-beta mRNA species in Northern blotting experiments, with maximal induction occurring 48 h after addition of 1,25-(OH)2D3. Analyses of 1,25-(OH)2D3-induced PKC mRNA expression by nuclear run-on transcription experiments suggest that the observed increases in PKC mRNA levels may occur by a posttranscriptional mechanism(s). In contrast to the transient increases in PKC mRNA levels, the increases in PKC Mr 80,000 protein species and in PKC enzyme activity were progressive in HL-60 cells treated with 1,25-(OH)2D3 between 1 and 5 days, thus implying the existence of a further up-regulation of PKC proteins occurring at the translational and/or posttranslational levels.(ABSTRACT TRUNCATED AT 250 WORDS)
人早幼粒细胞白血病细胞系HL-60在用各种诱导剂处理后可在体外分化。先前已表明，在二甲基亚砜或视黄酸诱导的HL-60细胞粒细胞分化过程中，蛋白激酶C（PKC）同工酶受到调节（M. Makowske、R. Ballester、Y. Cayre和O.M. Rosen，《生物化学杂志》，263: 3402 - 3410，1988；K. Hashimoto、A. Kishimoto、H. Aihara、I. Yasuda、K. Mikawa和Y. Nishizuka，《欧洲生物化学学会联合会快报》，263: 31 - 34，1990）。HL-60对1α,25 - 二羟基维生素D3（1,25-(OH)2D3）或12 - O - 十四烷酰佛波醇 - 13 - 乙酸酯有反应，可产生单核细胞。在本研究中，我们证明用1,25-(OH)2D3处理HL-60细胞会导致通过用PKC同工型特异性抗体进行免疫印迹检测到的PKC-α和PKC-β蛋白水平以及Ca(2+)和磷脂依赖性蛋白激酶活性显著增加。我们还在Northern印迹实验中观察到PKC-α和PKC-β mRNA种类的稳态水平短暂增加，在添加1,25-(OH)2D3后48小时出现最大诱导。通过核转录实验对1,25-(OH)2D3诱导的PKC mRNA表达进行分析表明，观察到的PKC mRNA水平增加可能通过转录后机制发生。与PKC mRNA水平的短暂增加相反，在用1,25-(OH)2D3处理1至5天的HL-60细胞中，PKC 80,000 Mr蛋白种类和PKC酶活性的增加是渐进的，因此意味着在翻译和/或翻译后水平存在PKC蛋白的进一步上调。（摘要截短于250字）
