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Objective To investigate the shRNA interference suppression Golgi targeting P115 gene expression in human gastric cancer cells BGC-823 invasion of , and possible molecular mechanisms involved . Methods pGPU6/GFP/Neo/P115-1318 stably transfected cancer cells BGC-823, inverted fluorescence microscope and calculate the transfection efficiency by Western-blot and RT-PCR analysis before and after transfection P115 protein and gene expression, evaluation inhibitory effect ; Western-blot and immunofluorescence chemical detection after transfection MIF, MMP2, MMP9 protein expression , RT-PCR detection of gene expression levels , cells in vitro wound healing assay and Transwell invasion assay after transfection BGC-823 cells metastasis abilities change . Results successfully pGPU6/GFP/Neo/P115-1318 stably transfected to gastric cancer BGC-823 cells , 48h after transfection average rate of about 68% ; cells with two control groups ( control group and shNC negative control group ) phase ratio , P115shRNA cells of P115, MIF, MMP2, MMP9 protein and gene expression levels were significantly lower (P lt; 0.05); cell scratch Transwell chamber experiments and in vitro invasion assay showed that cells compared with two control groups , stably transfected with the recombinant plasmid pGPU6/GFP/Neo/P115-1318 BGC-823 gastric cancer cell invasion cells also significantly reduced (P lt; 0.05). Conclusion BGC-823 human gastric cancer cells , the expression of P115 after interference suppression , possibly through reduced MIF, MMP2, MMP9 expression inhibits tumor cell invasion campaign.
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