Effect of ART1 gene silencing on metastatic potential of mouse colon cancer CT26 cells and its mechanism

Objective: To investigate the effect of ART1 (arginine-specific mono-ADP-ribosyltransferase-1) gene silencing on metastatic potential of mouse colon cancer CT26 cells, and to explore its possible mechanism. Methods: Lentivirus of ART1-shRNA was infected into CT26 cells and the infection efficiency was observed by fluorescence microscope. The expression levels of ART1 mRNA and protein were detected by RT-PCR and Western blotting, respectively. The invasion and adhesion potencies were observed by Transwell chamber and cell matrix adhesion assay, respectively. The activity levels of MMP-9 (matrix metalloproteinase-9) and MMP-2 were determined by gelatin zymography assay. The expression levels of ART1, RhoA (Ras homolog gene family, member A), ROCK1 (Rho-associated coiled-coil contanining kinase 1), MMP-9 and MMP-2 were detected by Western blotting. Results: The stable cell lines infected with lentivirus of ART1-shRNA were obtained successfully. After transfection of ART1-shRNA, the expression levels of ART1 mRNA and protein were decreased in CT26 cells. As compered with the blank control group, the inhibitory rates of invasion and cell matrix adhesion of the cells in ART1-shRNA group (39.63% and 40.70%) were sinificantly diseased (both P < 0.05). The expression levels of RhoA, ROCK1, MMP-2 and MMP-9 proteins and the activities of MMP-2 and MMP-9 proteins in ART1-shRNA group were decreased (P < 0.05). Conclusion: ART1 gene silencing in CT26 cells can inhibite the abilities of cell invasion and cell matrix adhesion, which may be associated with ART1 gene silencing and the blocking-activities of RhoA, ROCK1, MMP-9 and MMP-2. All of these suggest that ART1 gene probably plays a significant role in invasion and metastasis of colon cancer. DOI:10.3781/j.issn.1000-7431.2013.06.002