The effect of PKC phosphorylation sites mutation in JWA coding region on TPA-induced MCF-7 cell differentiation / 中华劳动卫生职业病杂志
Chinese Journal of Industrial Hygiene and Occupational Diseases
; (12): 398-401, 2007.
Article
en Zh
| WPRIM
| ID: wpr-321984
Biblioteca responsable:
WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To investigate the effect of PKC phosphorylation sites mutation in JWA coding region on TPA-induced MCF-7 cell differentiation.</p><p><b>METHODS</b>Site directed gene mutation was used to construct one or two PKC sites mutations in pEGFP-N1-JWA vectors, and transfected into MCF-7 cells by polyfect reagent, and cell differentiation was characterized by accumulation of lipid droplet as indicated by positive Oil-red-O staining of cells.</p><p><b>RESULTS</b>All these transfected cell lines, MCF-7-N1(transfected with pEGFP-N1 vector), MCF-7-JWA(transfected with pEGFP-N1-JWA vector), MCF-7-JWA-1(transfected with PKC site 1 mutation pEGFP-N1-JWA vector), MCF-7-JWA-2(transfected with PKC site 2 mutation pEGFP-N1-JWA vector), MCF-7-JWA-1+2 (transfected with both PKC site 1 and 2 mutation pEGFP-N1-JWA vector) were treated with 20 nmol/L TPA for 48 h, and the percentages of positive Oil-red-O staining of cells were 48%, 67%, 69%, 67% and 70% respectively. The percentages of cell differentiation in JWA containing vectors transfected cells treated with TPA were significantly higher those of MCF-7-N1 cells (vector only control). However there were no significant differences between mutated and unmutated cells.</p><p><b>CONCLUSION</b>JWA transfection enhanced MCF-7 cell differentiation induced by TPA significantly, and PKC sites mutation in JWA coding region has no obviously effect on TPA-induced MCF-7 cell differentiation.</p>
Texto completo:
1
Base de datos:
WPRIM
Asunto principal:
Farmacología
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Fosforilación
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Proteína Quinasa C
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Acetato de Tetradecanoilforbol
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Transfección
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Diferenciación Celular
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Mutagénesis Sitio-Dirigida
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Mutación Puntual
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Línea Celular Tumoral
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Vectores Genéticos
Límite:
Female
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Humans
Idioma:
Zh
Revista:
Chinese Journal of Industrial Hygiene and Occupational Diseases
Año:
2007
Tipo del documento:
Article