Proteolysis-inducing factor (pif): cloning, expression and biological activities (2003)
- Authors:
- Autor USP: BELIZARIO, JOSE ERNESTO - ICB
- Unidade: ICB
- Assunto: FARMACOLOGIA
- Language: Português
- Abstract: Objetivo: PIF is a 24 kDa sulfated glycoprotein capable of inducing muscle protein degradation in isolated soleous muscle preparation and weight loss during experimental cancer cachexia. Métodos e Resultados: PIF is a gene localized in chromosome 12 and codes to a typical secretory precursor protein of 11 kDa with two N- and two O-glycosylation sites. The goal of our study is the development of biological assays for detecting and analysis of biological effects of PIF. First, using specific primers, we identified PIF expression by RT-PCR in metastatic breast carcinoma and melanoma cell lines and tissues. Subsequently, we also detected PIF message in the LPS and H2O2-stimulated human monocytes/T/B cells (PMBC) from healthy donors with a highly similar sequence of PIF from other sources. Second, we cloned and expressed PIF protein using the vectors pAE, pGEX4T2 and pcDNA and the E.coli and mammalian cell expression systems. The pcDNA/PIF plasmid was used for transfection and selection (Geneticin) of B16-F10 melanoma clone that overexpress PIF mRNA and protein. Third, we injected intravenously 8 x 105 B16 melanoma cells from pDNA clone (control) and clone overexpressor of human PIF gene to C57BL/6 mice (27-30 g) and evaluated weight loss, survival, and lung metastasis over 21 days period after the cell inoculation. The mice bearing the B16/PIF tumor had a progressive loss of body weight and mortality rate, whereas B-16/pcDNA maintained body mass and survivedfrom the tumor growth. The lung metastasis was significantly increased in mice bearing the B16/PIF tumour. Conclusões: These data support the idea that PIF is a novel cytokine involved in both cancer cachexia and metastasis processes
- Imprenta:
- Publisher: Federação de Sociedades de Biologia Experimental
- Publisher place: Curitiba, Paraná
- Date published: 2003
- Source:
- Título do periódico: Resumos
- Conference titles: Reunião Anual da Federação de Sociedades de Biologia Experimental, FeSBE
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ABNT
GARAY-MALPARTIDA, Humberto Miguel et al. Proteolysis-inducing factor (pif): cloning, expression and biological activities. 2003, Anais.. Curitiba, Paraná: Federação de Sociedades de Biologia Experimental, 2003. . Acesso em: 23 abr. 2024. -
APA
Garay-Malpartida, H. M., Markovic, J., Alves, J., Pádua, J. B., Kashiabara, J. A., Moura, R., et al. (2003). Proteolysis-inducing factor (pif): cloning, expression and biological activities. In Resumos. Curitiba, Paraná: Federação de Sociedades de Biologia Experimental. -
NLM
Garay-Malpartida HM, Markovic J, Alves J, Pádua JB, Kashiabara JA, Moura R, Camargo A, Belizário JE. Proteolysis-inducing factor (pif): cloning, expression and biological activities. Resumos. 2003 ;[citado 2024 abr. 23 ] -
Vancouver
Garay-Malpartida HM, Markovic J, Alves J, Pádua JB, Kashiabara JA, Moura R, Camargo A, Belizário JE. Proteolysis-inducing factor (pif): cloning, expression and biological activities. Resumos. 2003 ;[citado 2024 abr. 23 ] - Análise da expressão do gene humano PIF/DCD em camundongos transgênicos
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