FAK-related non-kinase plasmid transfection inhibited hepatic stellate cells proliferation.
- Author:
Xiao-Xia HUO
1
;
Xiao-Lan ZHANG
;
Jian-Gang SHEN
;
Juan WEI
;
Yong-Qing DOU
Author Information
- Publication Type:Journal Article
- MeSH: Cell Line; Cell Proliferation; Fibronectins; Hepatic Stellate Cells; cytology; Humans; Phosphorylation; Plasmids; genetics; Protein-Tyrosine Kinases; genetics; Transfection
- From: Chinese Journal of Applied Physiology 2009;25(1):69-73
- CountryChina
- Language:Chinese
-
Abstract:
AIMTo observe the effect of FAK-related non-Kinase (FRNK) plasmid on hepatic stellate cell (HSC) proliferation stimulated by fibronectin (FN).
METHODSFRNK plasmid was transfected into HSC with transient liposomal transfection. The proteins of FRNK, FAK and p-FAK(Tyr397) were assayed by Western blotting analysis. The proliferation of HSC was evaluated by improved MTT assay, and cell cycle pattern was determined by flow cytometry (FCM).
RESULTS(1) The expression of FRNK protein increased after FRNK transfected HSC, and it was at 48 h that the expression of FRNK protein was the highest (P < 0.01). The protein level of FAK was no significant difference between before FRNK plasmid transfection and after transfection (P > 0.05). The expression of p-FAK(Tyr397) protein was down-regulated after FRNK had been transfected in HSC, (P < 0.01). (2) The HSC proliferation inhibition rates at 12 h, 24 h and 48 h after FRNK transfection were 20.07%, 26.16%, 29.77%, respectively (P < 0.01). (3) Compared with the non-FRNK plasmid group, the FRNK-transfected HSCs almost arrested in G0/G1 phase (71.4 +/- 2.81 vs 48.9 +/- 1.66, P < 0.01).
CONCLUSIONAfter FRNK were transfected successfully in HSCs using lipofectamine, the phosphorylation of FAK was inhibited. The HSC proliferation was restrained in a time-dependent manner and the HSC was arrested in G0/G1 phase.
