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Mouse Fyn induces pseudopodium formation in Chinese hamster ovary cells
Lei An1, Shengnan Liu, Wei Zhang
1College of Veterinary Medicine, Northwest A&F University, Yangling 712100, China.
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.
Insights
Overexpression of Fyn protein in Chinese hamster ovary (CHO) cells induced significant changes in cell morphology. This led to the formation of filopodia and lamellipodia, promoting cell movement.
Area of Science:
- Cell Biology
- Molecular Biology
- Biochemistry
Background:
- The Fyn tyrosine kinase plays a role in various cellular processes.
- Understanding Fyn's molecular mechanisms is crucial for cell biology research.
Purpose of the Study:
- To investigate the molecular mechanisms of Fyn's effects on cell morphology, pseudopodium dynamics, and migration.
- To analyze the impact of Fyn overexpression on Chinese hamster ovary (CHO) cell behavior.
Main Methods:
- Subcloning the Fyn gene into a pEGFP-N1 vector for transfection into CHO cells.
- Monitoring Fyn gene and protein expression using reverse transcription-PCR (RT-PCR) and Western blotting.
- Analyzing cell morphology and dynamics via 4',6-diamidino-2-phenylindole staining and time-lapse imaging.
Main Results:
- Transfection with pEGFP-N1-Fyn significantly altered CHO cell morphology.
- Fyn expression was confirmed at both mRNA and protein levels.
- Overexpression of Fyn induced the formation of filopodia and lamellipodia.
Conclusions:
- Mouse Fyn overexpression promotes filopodia and lamellipodia formation in CHO cells.
- Fyn plays a key role in regulating cell shape and promoting cell migration.
