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[The effects of TMSG-1 gene transfection on metastatic phenotype of pg cancer cells].

OBJECTIVE
To observe the relationship between TMSG-1 gene and tumor metastatic phenotype.


METHODS
TMSG-1 cDNA fragment which contained full length open reading frame of TMSG-1 gene was cloned into pcDNA3 plasmid to reconstruct sense and antisense eukaryotic expression plasmids of TMSG-1 gene containing neo selection marker. Both sense and antisense eukaryotic expression plasmids of TMSG-1 gene were transfected into the highly metastatic subclone PG-BE1 by LipofectAMINE method and the positive clones were selected by G418. RT-PCR was used to examine the expression level of the transfected gene and the changes of biological characteristics were checked by a series of in vitro and in vivo assays.


RESULTS
The results showed that higher expression levels of TMSG-1 gene in BE1-S cells (cells transfected by sense TMSG-1 cDNA) than the control BE1 cells and BE1-V cells (cells transfected by pcDNA3 plasmid). The expression levels of TMSG-1 gene in BE1-AS cells (cells transfected by antisense TMSG-1 cDNA) were lower than those of the control BE1 cells and BE1-V cells. Compared with the control BE1 cells and BE1-V cells, BE1-AS cells grew more rapidly, and produced more foci in soft agar. Although the BE1-S cells did not reveal significant different growth capacity, the infiltrating ability and colony formation potential of BE1-S cells were decreased, compared with the control BE1 cells and BE1-V cells. Flow cytometry showed higher percentage of BE1-S cells in G0G1 phase than that of BE1 cells, and the presence of apoptotic peak in BE1-S cells.


CONCLUSION
The results suggest TMSG-1 gene may represent a tumor metastasis suppressor gene.