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Nuclear localization of insulin-like growth factor binding protein-6 / 中国医学科学院学报
Acta Academiae Medicinae Sinicae ; (6): 735-739, 2009.
Artigo em Chinês | WPRIM | ID: wpr-301617
ABSTRACT
<p><b>OBJECTIVE</b>To study the nuclear localization of insulin-like growth factor binding protein-6(IGFBP-6) in PC-3M cells.</p><p><b>METHODS</b>The two fragments of the nuclear localization sequence (NLS)-deleted IGFBP-6 and the NLS-mutated IGFBP-6 were obtained by overlapping PCR, and then the fragment was inserted into a pEGFP-C1 vector. PC-3M cells were transfected with the expression constructs containing wild-type IGFBP-6 or the two mutants (pEGFP-C1-BP6 Delta NLS and pEGFP-C1-BP6-Mut), and the different distribution of the three EGFP-fusion proteins was observed by confocal laser microscope. The statistical analysis of the ratio of the nuclear fluorescence to the cytoplasmic fluorescence (Fn/c) was performed. Results Confocal microscopic images of transfected cells showed that the green fluorescence of EGFP-IGFBP-6 was concentrated mostly in the nuclei, whereas the control cells expressing EGFP showed green fluorescence distributed uniformly. The results of Fn/c from EGFP and EGFP-IGFBP-6 were significant different (P<0.05). The NLS-deleted IGFBP-6 completely eliminated nuclear accumulation of the green fluorescent signal; in contrast, nuclear accumulation was only slightly reduced for the NLS-mutated IGFBP-6; compared with wild-type IGFBP-6, both mutants were significantly different (P<0.05). Conclusions IGFBP-6 can be translocated to the nucleus in PC-3M cell that is mediated by a putative NLS sequence. Our study provides new evidence for further studies on the insulin-like growth factor-independent activity of IGFBP-6.</p>
Assuntos
Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Plasmídeos / Transfecção / Núcleo Celular / Proteína 6 de Ligação a Fator de Crescimento Semelhante à Insulina / Transporte Proteico / Linhagem Celular Tumoral / Vetores Genéticos / Genética / Metabolismo Limite: Humanos Idioma: Chinês Revista: Acta Academiae Medicinae Sinicae Ano de publicação: 2009 Tipo de documento: Artigo

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Texto completo: DisponíveL Índice: WPRIM (Pacífico Ocidental) Assunto principal: Plasmídeos / Transfecção / Núcleo Celular / Proteína 6 de Ligação a Fator de Crescimento Semelhante à Insulina / Transporte Proteico / Linhagem Celular Tumoral / Vetores Genéticos / Genética / Metabolismo Limite: Humanos Idioma: Chinês Revista: Acta Academiae Medicinae Sinicae Ano de publicação: 2009 Tipo de documento: Artigo