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首页> 外文期刊>American Journal of Physiology >PMA and staurosporine affect expression of the PCK gene in LLC-PK1-F+ cells.
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PMA and staurosporine affect expression of the PCK gene in LLC-PK1-F+ cells.

机译：PMA和星形孢菌素影响LLC-PK1-F +细胞中PCK基因的表达。

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摘要

The addition of phorbol 12-myristate 13-acetate (PMA) to renal LLC-PK1-F+ cells caused a rapid decrease in the level of phosphoenolpyruvate carboxykinase (PCK) mRNA and reversed the stimulatory effects of exposure to acidic medium (pH 6.9, 10 mM HCO-3) or cAMP. In contrast, prolonged treatment with PMA increased the levels of PCK mRNA. The two effects correlated with the membrane translocation and downregulation of the alpha-isozyme of protein kinase C and were blocked by pretreatment with specific inhibitors of protein kinase C. The rapid decrease in PCK mRNA caused by PMA occurred with a half-life (t1/2 = 1 h) that is significantly faster than that measured during recovery from acid medium or following inhibition of transcription (t1/2 = 4 h). The effect of PMA was reversed by staurosporine, which apparently acts by inhibiting a signaling pathway other than protein kinase C. Staurosporine had no effect on the half-life of the PCK mRNA, but it stimulated the activity of a chloramphenicol acetyltransferase gene that was driven by the initial 490 base pairs of the PCK promoter and transiently transfected into LLC-PK1-F+ cells. This effect was additive to that of cAMP, and neither stimulation was reversed by PMA. The stimulatory effect of staurosporine was mapped to the cAMP response element (CRE-1) and P3(II) element of the PCK promoter. The data indicate that, in LLC-PK1-F+ cells, activation of protein kinase C decreases the stability of the PCK mRNA, whereas transcription of the PCK gene may be suppressed by a kinase that is inhibited by staurosporine.
机译：向肾LLC-PK1-F +细胞中添加佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）导致磷酸烯醇丙酮酸羧激酶（PCK）mRNA水平迅速降低，并逆转了暴露于酸性介质（pH 6.9，10）的刺激作用mM HCO-3）或cAMP。相反，长时间使用PMA治疗会增加PCK mRNA的水平。这两种作用与蛋白激酶C的膜易位和α-同工酶的下调相关，并被蛋白激酶C的特异性抑制剂预处理所阻断。由PMA引起的PCK mRNA的快速下降发生了半衰期（t1 / 2 = 1 h）的速度明显快于从酸性培养基中恢复或转录抑制后的测量速度（t1 / 2 = 4 h）。星形孢菌素逆转了PMA的作用，星形孢菌素显然通过抑制蛋白激酶C以外的信号传导途径起作用。星形孢菌素对PCK mRNA的半衰期没有影响，但它刺激了氯霉素乙酰基转移酶基因的活性。通过最初的PCK启动子的490个碱基对被瞬时转染到LLC-PK1-F +细胞中。这种作用是cAMP的附加作用，PMA均不能逆转刺激。星形孢菌素的刺激作用被映射到PCK启动子的cAMP反应元件（CRE-1）和P3（II）元件。数据表明，在LLC-PK1-F +细胞中，蛋白激酶C的激活降低了PCK mRNA的稳定性，而PCK基因的转录可能受到星形孢菌素抑制的激酶的抑制。

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