Mechanism of novel vitamin K analog induced growth inhibition in human hepatoma cell line
Mechanism of novel vitamin K analog induced growth inhibition in human hepatoma cell line
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DOI：
10.1016/s0168-8278(00)00102-1
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发表时间：
2001-05-01
影响因子：
25.7
通讯作者：
Carr, BI
中科院分区：
文献类型：
--
作者：
Osada, S;Carr, BI
背景/目的：为了了解肝再生或肝癌细胞凋亡的机制，估计生长因子受体信号转导的转折点非常重要，由于2(2-羟乙基硫芳基)3-甲基-1,4-萘醌或CPD 5已被证明可介导Hep3B肝癌细胞中表皮生长因子-(EGF)受体的磷酸化，因此EGF和CPD 5介导的信号转导之间的差异是研究方法：采用Hoechst荧光法测定DNA含量。通过蛋白质印迹或二维电泳描述磷酸化蛋白。结果：CPD 5诱导的EGFR磷酸化可刺激Ras通路。然而，CPD 5 介导的细胞外信号调节激酶 (ERK) 磷酸化并未因 PD153035 上游激活的抑制而拮抗。 CPD 5 抑制细胞裂解物中的 ERK 去磷酸化，表明 CPD 5 引起的 ERK 磷酸化取决于激酶活性和磷酸酶抑制。二维电泳显示额外的磷酸化ERK斑点，U0126拮抗生长抑制和该斑点的出现，表明该斑点与CPD 5诱导的生长抑制密切相关。结论：EGFR通路的转折点与磷酸化ERK的表达水平密切相关。 ERK 磷酸化被认为在生长因子诱导的信号转导中发挥关键作用，(C) 2001 年欧洲肝脏研究协会。由 Elsevier Science B,V 出版，保留所有权利。
Background/Aims: To understand the mechanisms of liver regeneration or hepatoma apoptosis, it is important to estimate the turning point of the signal transduction by growth factor receptor, Since 2 (2-hydroxyethylsulfaryl) 3-methyl-1,4-naphthoquinone or CPD 5 has been shown to mediate the phosphorylation of epidermal growth factor -(EGF) receptor in Hep3B hepatoma cells, the differences between EGF and CPD 5-mediated signal transduction were studied.Methods: DNA content was measured by Hoechst fluorescent assay. Phosphorylated proteins were described with Western blots or two-dimensional electrophoresis,Results: CPD 5-induced EGFR phosphorylation was functional to stimulate Ras pathway. However, CPD 5-mediated extracellular signal-regulated kinase (ERK) phosphorylation was not antagonized by inhibition of upstream activation with PD153035. CPD 5 inhibited ERK dephosphorylation in cell lysate, suggesting that ERK phosphorylation by CPD 5 was depending on kinase activity and phosphatase inhibition. Two-dimensional electrophoresis showed extra phospho ERK spot, which was indicated to have close association with CPD 5-induced growth inhibition, since U0126 antagonized growth inhibition and appearance of this spot.Conclusions: The turning point of EGFR pathway was proved to have close association with the expressed level of phosphorylated ERK. ERK phosphorylation was suggested to play a critical role in growth factor-induced signal transduction, (C) 2001 European Association for the Study of the Liver. Published by Elsevier Science B,V, All rights reserved.