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[The effects of TMSG-1 gene transfection on metastatic phenotype of pg cancer cells]
Ba Bian1, Chunshu Ma, Jiangfeng You
1Department of Pathology, Peking University School of Basic Medical Sciences, Beijing 100083, China.
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.
Insights
The TMSG-1 gene may suppress tumor metastasis. Overexpressing TMSG-1 decreased cancer cell infiltration and colony formation, suggesting its role in inhibiting tumor spread.
Area of Science:
- Molecular Biology
- Oncology
- Gene Expression Analysis
Context:
- Tumor metastasis is a complex process involving multiple genetic factors.
- Understanding genes that regulate metastasis is crucial for developing targeted therapies.
Purpose:
- To investigate the role of the TMSG-1 gene in regulating tumor metastatic phenotype.
- To determine if TMSG-1 acts as a tumor suppressor gene.
Summary:
- The TMSG-1 gene was cloned into expression plasmids and transfected into highly metastatic PG-BE1 cells.
- Overexpression of TMSG-1 (BE1-S cells) reduced cell infiltration and colony formation, while antisense transfection (BE1-AS cells) increased these properties.
- Flow cytometry revealed increased G0G1 phase arrest and apoptosis in BE1-S cells.
Impact:
- TMSG-1 gene expression is inversely correlated with tumor metastatic potential.
- TMSG-1 may function as a tumor metastasis suppressor gene.
- These findings could inform novel therapeutic strategies targeting TMSG-1 for cancer treatment.
