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佛波酯对A-549细胞株中蛋白激酶C的调节

Regulation of Protein Kinase C in A-549 Cells by Phorbo l Ester

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【作者】 陈军松柴敏强宋建国

【Author】 CHEN Jun-Song, CHAI Min Qiang, SONG Jian Guo * (State Key Laboratory of Molecular Biology, Shanghai Institute of Bioch emistry, Shanghai Institutes for Biological Sciences, the Chinese Academy of Sciences, Shanghai 200031, Chin a)

【机构】 中国科学院上海生命科学研究院中国科学院上海生物化学研究所分子生物学国家重点实验室!上海200031中国科学院上海生物化学研究所分子生物学国

【摘要】 探讨了佛波酯 ( phorbol 12 myristate 13 acetate,PMA)对人肺癌表皮细胞株A 5 4 9细胞中数种蛋白激酶C(proteinkinaseC ,PKC)亚型的调节作用。用蛋白质免疫印迹法在A 5 4 9细胞中检测到有PKC α、PKC βII、PKC γ、PKC δ和PKC ε等亚型的表达 ,但未检测到PKC ζ的表达。PMA对细胞的短时间处理诱导所有这 5种亚型的不同程度的转位 (即活化 )。长时间处理则显著下调PKC α、PKC γ、PKC δ和PKC ε的蛋白质水平 ,但PKC βII的蛋白质水平却不受影响。此外 ,PMA长时间处理细胞对同一PKC亚型在膜组分和胞液组分中的蛋白质水平表现出不同的调节作用。PMA长时间处理导致胞液组分中的PKC δ和PKC γ水平显著的下降 ,并引起PKC α和PKC βII的消失或耗空 ;然而 ,对细胞膜组分中这些PKC亚型的蛋白质水平却没有明显的影响。PMA长时间处理对胞液和胞膜组分中的PKC ε水平的下调作用均十分明显。以上的观察为PMA在活化与下调不同PKC亚型中的作用提供了新的证据 ,有助于增加对细胞内PKC的活化及水平的变化在化学致癌过程中的作用的了解

【Abstract】 Protein kinase C, a family of serine threonine pro te in kinases, mediates a variety of intracellular signaling events. Here, the regu latory effect of phorbol 12 myristate 13 acetate(PMA)on the several PKC isozym es in the human lung carcinoma cells A 549 was studied. The expression of PKC α?PKC βII?PKC γ?PKC δ and PKC ε in A 549 cells was examined. No det ectable PKC ζ was observed. Short term treatment of cells with PMA led to the translocation of these PKC isozymes, to different extent, from cytosol to cell membrane. Whereas, prolonged treatment of cells with PMA pronouncedly reduced th e levels of PKC α?PKC γ?PKC δ and PKC ε, but the intracellular level o f PKC βII was not affected. Furthermore, PMA was observed to have differential effects on the down regulation of PKC isozymes located in the cytosol and of t hose located in the membrane. Prolonged PMA treatment caused extensive decrease in the levels of cytosolic PKC δ and PKC γ, and depleted cytosolic PKC α a nd PKC βII. However, the amount levels of membrane PKC α?PKC βII?PKC γ ?PKC δ isozymes were not decreased. In contrast, PKC ε in both cytosol and membrane fraction was obviously down regulated by prolonged PMA treatment. This study provided novel evidence on the PMA mediated activation and down regulat ion of different PKC isozymes, which might be helpful in deepening our understan ding on the roles of PKC activation and the alterations of their intracellular l evels in processes of chemical carcinogenesis.

【关键词】 蛋白激酶C佛波酯转位蛋白质印迹A-549
【Key words】 protein kinase CPMAtranslocationWestern blot tingA 549
【基金】 国家自然科学基金资助项目 (No .3962 5 0 0 0 7;No .39870 396)~~
  • 【文献出处】 生物化学与生物物理学报 ,Acta Biochimica Et Biophysica Sinica , 编辑部邮箱 ,2000年06期
  • 【分类号】Q555.7
  • 【被引频次】6
  • 【下载频次】134
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