Mouse Fyn induces pseudopodium formation in Chinese hamster ovary cells.
10.4142/jvs.2014.15.1.111
- Author:
Lei AN
1
;
Shengnan LIU
;
Wei ZHANG
;
Yamei ZHANG
;
Yingxue HUANG
;
Xinde HU
;
Shulin CHEN
;
Shanting ZHAO
Author Information
1. College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China. shantingzhao@hotmail.com
- Publication Type:Original Article ; Research Support, Non-U.S. Gov't
- Keywords:
filopodia;
Fyn;
lamellipodia;
time-lapse
- MeSH:
Animals;
Blotting, Western;
CHO Cells;
Cricetinae;
Cricetulus;
Genetic Vectors;
Green Fluorescent Proteins/genetics;
Mice;
Proto-Oncogene Proteins c-fyn/genetics/*metabolism;
Pseudopodia/*metabolism;
Reverse Transcriptase Polymerase Chain Reaction;
Time-Lapse Imaging;
Transfection
- From:Journal of Veterinary Science
2014;15(1):111-115
- CountryRepublic of Korea
- Language:English
-
Abstract:
Molecular mechanisms underlying the effects of Fyn on cell morphology, pseudopodium movement, and cell migration were investigated. The Fyn gene was subcloned into pEGFP-N1 to produce pEGFP-N1-Fyn. Chinese hamster ovary (CHO) cells were transfected with pEGFP-N1-Fyn. The expression of Fyn mRNA and proteins was monitored by reverse transcription-PCR and Western blotting. Additionally, transfected cells were stained with 4',6-diamidino-2-phenylindole and a series of time-lapse images was taken. Sequences of the recombinant plasmids pMD18-T-Fyn and pEGFP-N1-Fyn were confirmed by sequence identification using National Center for Biotechnology Information in USA, and Fyn expression was detected by RT-PCR and Western blotting. The morphology of CHO cells transfected with the recombinant vector was significantly altered. Fyn expression induced filopodia and lamellipodia formation. Based on these results, we concluded that overexpression of mouse Fyn induces the formation of filopodia and lamellipodia in CHO cells, and promotes cell movement.
