磷脂酰胆碱合成的酚酯刺激需要蛋白激酶C-α和磷脂酶D的表达

Zoltan Kiss, Karan S. Crilly, Wayne H. Anderson
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引用次数: 8

摘要

蛋白激酶C (PKC)激活剂phorbol 12-肉豆蔻酸酯13-乙酸酯(PMA)刺激磷脂酶D (PLD)介导的磷脂酰胆碱(PtdCho)的合成和水解。本研究利用附着和悬浮的NIH 3T3成纤维细胞以及表达PKC-α和不同水平PtdCho特异性PLD活性的MCF-7人乳腺癌细胞系的变异体来确定PKC-α、PtdCho水解和胆碱摄取在PMA对PtdCho合成的影响中的可能作用。在PKC-α和PLD活性均极低表达的野生型MCF-7细胞中,PMA对[14C]胆碱摄取或掺入PtdCho的影响很小。在高表达PKC-α但缺乏PtdCho特异性PLD活性的多药耐药MCF-7/MDR1细胞中,100 nm PMA对[14C]胆碱的摄取(约1.5倍)和[14C]PtdCho的合成(1.5至2倍)具有相对较小的刺激作用。在高水平表达PKC-α和PLD活性的NIH 3T3成纤维细胞和MCF-7/PKC-α细胞中,10-100-nM PMA仅略微增强[14C]胆碱摄取(1.7- 2.2倍),而对PtdCho合成具有更大的(~ 4 - 9倍)刺激作用。PMA在MCF-7/PKC-α细胞中显著促进磷脂酸(PtdOH)的形成(增加2.8倍),但在MCF-7/MDR1细胞中没有(增加1.4倍),而在两种细胞系中,PMA对1,2-二酰基甘油(1,2- dag)的形成只有很小的刺激作用(1.3 - 1.5倍)。在悬浮的NIH 3T3细胞中,200-300-mM乙醇阻断了PMA对PtdOH形成的刺激作用,但不影响PtdCho的合成,这表明PtdOH及其衍生的1,2- dag都不是PMA对PtdCho合成作用的中介。在附着的NIH 3T3细胞中,二甲基苯[a]蒽增强了磷脂胆碱的形成,从而增强了胆碱的摄取,而没有增加PtdCho的合成或改变PMA的作用。虽然结果表明PMA对PtdCho合成的刺激作用需要PKC-α和PtdCho特异性PLD的表达,但它们不支持1,2- dag、PtdOH或胆碱在PMA作用中的中介作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Phorbol ester stimulation of phosphatidylcholine synthesis requires expression of both protein kinase C-α and phospholipase D

The protein kinase C (PKC) activator phorbol 12-myristate 13-acetate (PMA) stimulates both the synthesis and phospholipase D (PLD)-mediated hydrolysis of phosphatidylcholine (PtdCho). Here, attached and suspended NIH 3T3 fibroblasts as well as variants of the MCF-7 human breast carcinoma cell line expressing PKC-α and a PtdCho-specific PLD activity at widely different levels were used to determine the possible role of PKC-α, PtdCho hydrolysis, and choline uptake in the mediation of PMA effect on PtdCho synthesis. In wild-type MCF-7 cells, which express both PKC-α and PLD activities at very low levels, PMA had little effects on the uptake or incorporation [14C]choline into PtdCho. In multidrug resistant MCF-7/MDR1 cells, which highly express PKC-α but lack the PtdCho-specific PLD activity, 100-nM PMA had relatively small stimulatory effects on the uptake of [14C]choline (∼1.5-fold) and [14C]PtdCho synthesis (1.5- to 2-fold). In NIH 3T3 fibroblasts and MCF-7/PKC-α cells, both expressing PKC-α and PLD activities at high levels, 10–100-nM PMA enhanced [14C]choline uptake only slightly (1.7- to 2.2-fold), while it had much greater (∼4–9-fold) stimulatory effects on PtdCho synthesis. PMA significantly enhanced the formation of phosphatidic acid (PtdOH) in MCF-7/PKC-α cells (2.8-fold increase), but not in MCF-7/MDR1 cells (1.4-fold increase), while in both cell lines it had only small (1.3–1.5-fold) stimulatory effects on 1,2-diacylglycerol (1,2-DAG) formation. In suspended NIH 3T3 cells, 200–300-mM ethanol blocked the stimulatory effect of PMA on PtdOH formation without affecting PtdCho synthesis indicating that neither PtdOH nor 1,2-DAG derived from it is a mediator of PMA effect on PtdCho synthesis. In attached NIH 3T3 cells, dimethylbenz[a]anthracene enhanced phosphocholine formation and, thus, choline uptake without increasing PtdCho synthesis or modifying the effect of PMA. While the results indicate that the stimulatory effect of PMA on PtdCho synthesis requires the expression of both PKC-α and a PtdCho-specific PLD, they do not support a role for 1,2-DAG, PtdOH or choline in the mediation of PMA effect.

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