Sustained small and intermediate size proteins expression in phorbol 12-myristate 13-acetate/ionomycine prolonged stimulated human fibroblasts
Asian Pacific Journal of Tropical Biomedicine
; (12): 432-436, 2017.
Article
en Zh
| WPRIM
| ID: wpr-609701
Biblioteca responsable:
WPRO
ABSTRACT
Objective:
To compare the protein profile of culture supematants in stimulated and unstimulated human fibroblasts to find some proteins indicating the presence of fibroblasts and their activation status.Methods:
Dcrmal fibroblasts were stimulated with phorbol 12-myristate 13-acetate (PMA)/ionomycine for 72 h.MTT assay was done to determine cell viability and A/E fluorescent staining was used to evaluate the cell death pattern.Protein analysis was performed by gradient SDS polyacrylamide gel electrophoresis 8%-16%.Results:
The supernatant of 24 h cultured both stimulated and unstimulated fibroblasts showed two bands in SDS-PAGE analysis with relative molecular weights of 8.59 and 78,8 kDa.These bands density was decreased during the next 48 h in unstimulated cells while their expression was continued in PMA or PMA/ionomycine stimulated cells and a new 85.3 kDa band was appeared in unstimulated and 72 h PMA stimulated cells.Moreover,we found another seven small size (10-19.5 kDa) proteins in supernatants of 48 h and 72 h unstimulated but not in PMA or PMA/lonomycine stimulated fibroblasts.Most of these proteins expression were down regulated following fibroblast activation.This down-regulation is consistent with our finding that PMA or PMA/ionomycine stimulated cells exhibited a significant level of apoptosis cell death.ConcLusions:
Human fibroblasts produce some small to intermediate sized proteins with specific SDS-PAGE profile upon cell activation.Most of these proteins can be excreted in urine and can be immunogen theoretically so this data provided a reliable clue for fibrosis biomarker screening based on designation of an appropriated immunoassay.
Texto completo:
1
Base de datos:
WPRIM
Idioma:
Zh
Revista:
Asian Pacific Journal of Tropical Biomedicine
Año:
2017
Tipo del documento:
Article