Effect of up-regulation of Tspan-5 expression on the metastatic ability of colorectal cancer cells / 解放军医学杂志

ABSTRACT

Objective To investigate the effect of up-regulation of tetraspanin-5 (Tspan-5) expression on the metastatic ability of colorectal cancer (CRC) cells. Methods The coding sequence of Tspan-5 mRNA was amplified from total RNA of LoVo cells by RT-PCR and cloned into pEGFP-C1 to construct Tspan-5/pEGFP-C1,a eukaryotic expression vector. The expression of Tspan-5 in LST-R1 cells was up-regulated by gene tranfection. Western blotting was used to detect the expression level of Tspan-5/EGFP fusion protein,and laser scanning confocal microscopy (LSCM) was used to observe the distribution of Tspan-5/EGFP in LST-R1 cells. The changes in adhesion,migration and invasion ability of LST-R1 cells were determined by adhesion,migration and invasion experiment in vitro,respectively. Liver metastasis model of nude mice was used to test the changes in liver metastasis ability of LST-R1 cells in vivo. Results The Tspan-5/EGFP fusion protein could be detected by Western blotting in Tspan-5/pEGFP-C1 transfectants. LSCM showed that Tspan-5/EGFP protein located in cell membrane. Adhesion and migration assays showed that up-regulation of Tspan-5 expression level significantly promoted the adhesion and migration ability of LST-R1 cells on a series of extracellular matrix (ECM) components,including collagen IV,FN,LN and VN (P