Construction of eukaryotic expression plasmid for Rac1 gene and its effect on the biological function of colon cancer cells

doi:10.3877/cma.j.issn.1674-0793.2014.03.003

Abstract

Abstract:

Objective

To reveal the role of Rac1 gene regulating colon cancer cell invasion and migration.

Methods

Rac1 gene was amplified from colon cancer cells, and then constructed to pCDNA3.1(+)-Rac1 and pEGFP-Rac1 eukaryotic expression plasmid for overexpressed Rac1 protein. Meanwhile, siRNA was used to inhibit Rac1 gene expression. After transfecting Rac1-plasmid or siRNA into cells, Matrigel invasion, wound healing and Pull-down assays were performed to investigate the alteration of invasion, migration and Rac1 GTPase activity in colon cancer cells, respectively.

Results

A eukaryotic expression plasmid containing Rac1 gene was successfully constructed. Up or down-regulation of Rac1 expression could stimulate or block GTPase activation, and Rac1 gene had the ability to modulate the migration and invasion of colon cancer cells.

Conclusions

Our study provides a potential target that gene Rac1 plays the synergistic role with conventional therapy in colon cancer by suppressing Rac1 expression.

Key words: Colon cancer, Rac1 gene, Eukaryotic expression plasmid, GTPase

Zhiqiang Ye, Yi Chen, Yaqiong Chen, Bo Liu. Construction of eukaryotic expression plasmid for Rac1 gene and its effect on the biological function of colon cancer cells[J]. Chinese Archives of General Surgery(Electronic Edition), 2014, 08(03): 185-190.

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URL: https://zhptwkxwx.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-0793.2014.03.003

https://zhptwkxwx.cma-cmc.com.cn/EN/Y2014/V08/I03/185

Figures/Tables 8

References 16

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