Phorbol ester stimulation of phosphatidylcholine synthesis requires expression of both protein kinase C-α and phospholipase D

Zoltan Kiss, Karan S. Crilly, Wayne H. Anderson
{"title":"Phorbol ester stimulation of phosphatidylcholine synthesis requires expression of both protein kinase C-α and phospholipase D","authors":"Zoltan Kiss,&nbsp;Karan S. Crilly,&nbsp;Wayne H. Anderson","doi":"10.1016/S0005-2760(98)00030-7","DOIUrl":null,"url":null,"abstract":"<div><p>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-<em>α</em> and a PtdCho-specific PLD activity at widely different levels were used to determine the possible role of PKC-<em>α</em>, PtdCho hydrolysis, and choline uptake in the mediation of PMA effect on PtdCho synthesis. In wild-type MCF-7 cells, which express both PKC-<em>α</em> and PLD activities at very low levels, PMA had little effects on the uptake or incorporation [<span><math><msup><mi></mi><mn>14</mn></msup><mtext>C</mtext></math></span>]choline into PtdCho. In multidrug resistant MCF-7/MDR1 cells, which highly express PKC-<em>α</em> but lack the PtdCho-specific PLD activity, 100-nM PMA had relatively small stimulatory effects on the uptake of [<span><math><msup><mi></mi><mn>14</mn></msup><mtext>C</mtext></math></span>]choline (∼1.5-fold) and [<span><math><msup><mi></mi><mn>14</mn></msup><mtext>C</mtext></math></span>]PtdCho synthesis (1.5- to 2-fold). In NIH 3T3 fibroblasts and MCF-7/PKC-<em>α</em> cells, both expressing PKC-<em>α</em> and PLD activities at high levels, 10–100-nM PMA enhanced [<span><math><msup><mi></mi><mn>14</mn></msup><mtext>C</mtext></math></span>]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-<em>α</em> 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-<em>α</em> and a PtdCho-specific PLD, they do not support a role for 1,2-DAG, PtdOH or choline in the mediation of PMA effect.</p></div>","PeriodicalId":100162,"journal":{"name":"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"1998-05-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1016/S0005-2760(98)00030-7","citationCount":"8","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Biochimica et Biophysica Acta (BBA) - Lipids and Lipid Metabolism","FirstCategoryId":"1085","ListUrlMain":"https://www.sciencedirect.com/science/article/pii/S0005276098000307","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 8

Abstract

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.

查看原文
分享 分享
微信好友 朋友圈 QQ好友 复制链接
本刊更多论文
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
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。
求助全文
约1分钟内获得全文 去求助
来源期刊
自引率
0.00%
发文量
0
期刊最新文献
Effect of methylglyoxal on the physico-chemical and biological properties of low-density lipoprotein Cholesterol synthesis is increased in mixed hyperlipidaemia The level of 7-dehydrocholesterol in plasma reflects the activity of 3-hydroxy-3-methylglutaryl coenzyme A reductase in the human liver Fatty acid α-oxidation of tetradecylthioacetic acid and tetradecylthiopropionic acid in cucumber (Cucumis sativus) Inductive electron-withdrawal from ammonium ion headgroups of cationic lipids and the influence on DNA transfection
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
现在去查看 取消
×
提示
确定
0
微信
客服QQ
Book学术公众号 扫码关注我们
反馈
×
意见反馈
请填写您的意见或建议
请填写您的手机或邮箱
已复制链接
已复制链接
快去分享给好友吧!
我知道了
×
扫码分享
扫码分享
Book学术官方微信
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术
文献互助 智能选刊 最新文献 互助须知 联系我们:info@booksci.cn
Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。
Copyright © 2023 Book学术 All rights reserved.
ghs 京公网安备 11010802042870号 京ICP备2023020795号-1