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首页> 外文期刊>Cellular Physiology and Biochemistry >GH3B6 Pituitary Tumor Cell Proliferation is Mediated by PKCα and PKCε via ERK 1/2-dependent Pathway
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GH3B6 Pituitary Tumor Cell Proliferation is Mediated by PKCα and PKCε via ERK 1/2-dependent Pathway

机译：GH3B6垂体肿瘤细胞增殖是通过ERK 1/2依赖性途径由PKCα和PKCε介导的

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Background In this report, we explored the role of PKCα and PKCε as mediators of phorbol 12-myristate13-acetate (PMA)-induced proliferation in pituitary tumor GH3B6 cells, and determined if the ERK1/2 and Akt pathways were activated. Methods The GH3B6 cell proliferation was estimated by BrdU incorporation and the cell cycle progression by flow cytometric cell cycle analysis. We determined the expression of PKCα and PKCε in membrane and cytosolic fractions by western blotting. The subcellular redistribution of both PKC isozymes was analyzed by confocal microscopy. Results Incubation with PMA for 15 min stimulated PKCα and PKCε activation, which was correlated with the phosphorylation of ERK1/2 but not Akt. The activation of both these PKC isozymes was closely associated with the stimulation of proliferation and the cell cycle progression induced by PMA in GH3B6 cells, an effect that was blocked by the inhibitors of PKCα (Gö6976) and PKCε (εV1-2). In addition, the pretreatment with the inhibitor of ERK1/2 (PD98059) prevented the mitogenic activity induced by treatment with PMA for 15 min. Conclusion We demonstrated that the activation of PKCα and PKCε by phorbol ester in tumor pituitary GH3B6 cells led to cell proliferation and cell cycle progression, effects that involved ERK1/2 activation.
机译：背景技术在本报告中，我们探讨了PKCα和PKCε作为佛波醇12-肉豆蔻酸13-乙酸酯（PMA）诱导的垂体瘤GH3B6细胞增殖的介质，并确定ERK1 / 2和Akt途径是否被激活。方法通过BrdU掺入来评估GH3B6细胞的增殖，并通过流式细胞术分析细胞周期。我们通过蛋白质印迹法确定了PKCα和PKCε在膜和胞质组分中的表达。通过共聚焦显微镜分析了两种PKC同工酶的亚细胞再分布。结果PMA孵育15分钟可刺激PKCα和PKCε活化，这与ERK1 / 2的磷酸化有关，而与Akt无关。这两种PKC同工酶的激活与GH3B6细胞中PMA诱导的增殖刺激和细胞周期进程密切相关，这一作用被PKCα（Gö6976）和PKCε（εV1-2）的抑制剂所阻断。此外，用ERK1 / 2抑制剂（PD98059）进行的预处理可防止PMA处理15分钟诱导的有丝分裂活性。结论我们证明佛波酯在肿瘤垂体GH3B6细胞中激活PKCα和PKCε导致细胞增殖和细胞周期进程，涉及ERK1 / 2激活。

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