Suppressing migration and invasion potencies of tongue carcinoma cells by lentiviral victor mediated shRNA specially silencing of phosphatase of regenerating liver cell-1 gene expression

Abstract:

Objective：To study the effect of specially silencing of phosphatase of regenerating liver cell-1（PRL-1） gene expression on migration and invasion potencies of tongue carcinoma cells. Methods：Five interfering sequences of shRNA targeting PRL-1 gene were designed，synthesized and constructed in corresponding plasmids and were confirmed by PCR electrophoresis and DNA sequencing. The most effective sequence of shRNA was screened by Western blot and the lentivirus was produced and packaged. TCA8113 cells were infected with recombinant lentivirus and the stably transfected cells were filtered and obtained. The silencing efficiency of PRL-1 gene was assessed by real-time PCR. The migration and invasion potencies of PRL-1 gene silenced cells were analyzed by wound healing and Transwell invasion assay. Results：PCR electrophoresis and DNA sequencing revealed that 5 shRNA sequences were di－rectionally constructed in plasmids. PRL-1-shRNA-1 was the most effective interference sequence selected by Western blot. The sta－bly infected TCA8113 cells were established after interfering and filtering. The expression of PRL-1 gene mRNA was decreased sig－nificantly（F=809.120，P=0.000）. After silencing the PRL-1 gene expression，the migration and invasion potencies of TCA8113 cells were significantly suppressed（F=1 092.970，P=0.000）. Conclusion：Silencing of PRL-1 gene expression can inhibit the migration and invasion potencies of TCA8113 cells significantly.