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首页> 外文期刊>Cytokine >Evidence for a novel thrombopoietin signalling event: activation of protein kinase A in human megakaryoblastic CMK cells.
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Evidence for a novel thrombopoietin signalling event: activation of protein kinase A in human megakaryoblastic CMK cells.

机译:新型血小板生成素信号转导事件的证据:人巨核细胞CMK细胞中蛋白激酶A的激活。

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摘要

Thrombopoietin (TPO) plays a crucial role in megakaryocyte development. TPO signalling, which is mediated by its receptor Mpl, includes Janus kinase, (JAK) signal transducer and activator of transcription (STAT) and Shc/Ras/mitogen-activated protein kinase (MAPK) pathways. The precise nature of these signalling routes has not been clarified in detail up until now. We investigated the effect of TPO on activation of cAMP-dependent protein kinase (PKA) and its involvement in MAPK signalling in human megakaryoblastic leukaemia CMK cells. For estimation of PKA activity, phosphorylation of a PKA-specific peptide substrate was assayed in CMK cell lysates. Since activation of PKA is associated with translocation of its catalytic subunit alpha (C-PKA) into the cell nucleus, Western blot analysis of nuclear fractions with an anti-C-PKA antibody was additionally performed. The activation of TPO-induced MAPK activation and the effect of the PKA inhibitor H-89 was measured using immunoblotting with a monoclonal anti-pERK antibody. TPO enhanced cAMP and induced activation of PKA in CMK cells. In addition, H-89 partly blocked TPO-induced MAPK activation in CMK cells. Our results indicate a novel TPO-triggered signalling event, activation of the cAMP/PKA pathway in human megakaryoblastic CMK cells. This signal transduction route seems to be involved in TPO-induced MAPK signaling.
机译:血小板生成素(TPO)在巨核细胞发育中起关键作用。由其受体Mpl介导的TPO信号传导包括Janus激酶(JAK)信号转导子和转录激活子(STAT)和Shc / Ras /丝裂原激活的蛋白激酶(MAPK)途径。到目前为止,尚未详细阐明这些信号传递途径的确切性质。我们调查了TPO对cAMP依赖性蛋白激酶(PKA)活化及其在人巨核细胞白血病CMK细胞中MAPK信号传导中的作用。为了估计PKA活性,在CMK细胞裂解物中测定了PKA特异性肽底物的磷酸化。由于PKA的激活与其催化亚基α(C-PKA)易位到细胞核有关,因此需要另外用抗C-PKA抗体进行Western印迹分析。使用单克隆抗pERK抗体的免疫印迹法测量了TPO诱导的MAPK活化的激活和PKA抑制剂H-89的作用。 TPO增强了cAMP并诱导了CMK细胞中PKA的激活。此外,H-89部分阻断了CMK细胞中TPO诱导的MAPK活化。我们的结果表明,一种新型的TPO触发的信号事件,即人类巨核细胞CMK细胞中cAMP / PKA途径的激活。该信号转导途径似乎与TPO诱导的MAPK信号传导有关。

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