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Proteomic analysis of novel targets associated with the enhancement of TrkA-induced SK-N-MC cancer cell death caused by NGF
Experimental & Molecular Medicine ; : e235-2016.
Article in English | WPRIM | ID: wpr-25937
ABSTRACT
Nerve growth factor (NGF) is known to regulate both cancer cell survival and death signaling, depending on the cellular circumstances, in various cell types. In this study, we showed that NGF strongly upregulated the protein level of tropomyosin-related kinase A (TrkA) in TrkA-inducible SK-N-MC cancer cells, resulting in increases in various TrkA-dependent cellular processes, including the phosphorylation of c-Jun N-terminal kinase (JNK) and caspase-8 cleavage. In addition, NGF enhanced TrkA-induced morphological changes and cell death, and this effect was significantly suppressed by the JNK inhibitor SP600125, but not by the phosphatidylinositol 3-kinase (PI3K) inhibitor wortmannin. To investigate novel targets associated with the enhancement of TrkA-induced SK-N-MC cell death caused by NGF, we performed Coomassie Brilliant Blue staining and two-dimensional (2D) proteomic analysis in TrkA-inducible SK-N-MC cells. We identified 31 protein spots that were either greatly upregulated or downregulated by TrkA during NGF treatment using matrix-associated laser desorption/ionization time of flight/time of flight mass spectrometry, and we analyzed the effects of SP600125 and wortmannin on the spots. Interestingly, 11 protein spots, including heterogeneous nuclear ribonucleoprotein K (hnRNP K), lamin B1 and TAR DNA-binding protein (TDP43), were significantly influenced by SP600125, but not by wortmannin. Moreover, the NGF/TrkA-dependent inhibition of cell viability was significantly enhanced by knockdown of hnRNP K using small interfering RNA, demonstrating that hnRNP K is a novel target associated with the regulation of TrkA-dependent SK-N-MC cancer cell death enhanced by NGF.
Subject(s)
Full text: Available Index: WPRIM (Western Pacific) Main subject: Phosphorylation / Phosphotransferases / Mass Spectrometry / Cell Survival / Cell Death / Nerve Growth Factor / Heterogeneous-Nuclear Ribonucleoprotein K / RNA, Small Interfering / JNK Mitogen-Activated Protein Kinases / Caspase 8 Type of study: Prognostic study Language: English Journal: Experimental & Molecular Medicine Year: 2016 Type: Article

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Full text: Available Index: WPRIM (Western Pacific) Main subject: Phosphorylation / Phosphotransferases / Mass Spectrometry / Cell Survival / Cell Death / Nerve Growth Factor / Heterogeneous-Nuclear Ribonucleoprotein K / RNA, Small Interfering / JNK Mitogen-Activated Protein Kinases / Caspase 8 Type of study: Prognostic study Language: English Journal: Experimental & Molecular Medicine Year: 2016 Type: Article