The effect of PKC phosphorylation sites mutation in JWA coding region on TPA-induced MCF-7 cell differentiation.
- Author:
Jian YE
1
;
Ai-ping LI
;
Wen QIU
;
Jin-hong ZHOU
;
Jian-wei ZHOU
Author Information
- Publication Type:Journal Article
- MeSH: Cell Differentiation; drug effects; Cell Line, Tumor; Female; Genetic Vectors; Humans; Mutagenesis, Site-Directed; Phosphorylation; Point Mutation; Protein Kinase C; genetics; metabolism; Tetradecanoylphorbol Acetate; pharmacology; Transfection
- From: Chinese Journal of Industrial Hygiene and Occupational Diseases 2007;25(7):398-401
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of PKC phosphorylation sites mutation in JWA coding region on TPA-induced MCF-7 cell differentiation.
METHODSSite 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.
RESULTSAll 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.
CONCLUSIONJWA 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.
