目的：通过uPAR特异siRNA靶向下调大肠癌LOVO细胞uPAR的表达，研究大肠癌细胞恶性生物学行为的变化。方法：构建3对uPAR特异siRNA，将其转入大肠癌LOVO细胞，用Western Blot检测uPAR-siRNA干扰效果，选择干扰效率最高的一对siRNA进行后续实验。利用流式细胞术及CCK-8试验检测uPAR-siRNA转染后大肠癌细胞增殖变化，利用Transwell小室实验及划痕实验检测uPAR-siRNA转染后对大肠癌细胞迁移能力变化。结果：大肠癌细胞分别转染3对uPAR-siRNA后，uPAR的表达明显下降，其中uPAR-siRNA-3的干扰效率最高；大肠癌细胞转染uPAR-siRNA-3后，G1期细胞数量明显增加，细胞增殖能力明显下降，细胞迁移数量明显减少及细胞迁移距离明显下降(均P<0.05)。结论：uPAR特异siRNA转染大肠癌细胞LOVO后，明显下调uPAR的表达；uPAR-siRNA转染大肠癌细胞LOVO后，明显抑制大肠癌细胞的增殖和迁移能力。%Objective: To investigate the change of malignant behaviors in colorectal cancer cells(LOVO) through targeted down-regulation the expression of uPAR by the specific uPAR-siRNA. Methods: Three pairs of specific uPAR-siRNA were constructed and transfected into LOVO colorectal cancer cell line. The interference efficiencies of uPAR-siRNAs were detected by Western Blot. The uPAR-siRNA with the highest interference efficiency was selected for the following experiments. The proliferation of colorectal cancer cells after transfection with uPAR-siRNA was detected by flow cytometry and CCK-8 assay. The migration of colorectal cancer cells after transfection with uPAR-siRNA was determinated by Transwell chamber system and Wound healing assay. Results: The expressions of uPAR in the colorectal cancer cells were significantly down-regulated after transfection with uPAR-siRNAs, and uPAR-siRNA-3 caused the most reduction of uPAR. After transfection with uPAR-siRNA-3 into colorectal cancer cells, the number in G1 phase was significantly increased, but the abilities of proliferation and migration were significantly inhibited , compared with the control group (all P<0.05). Conclusions: The expression of uPAR in colorectal cancer cells after transfection with the specific siRNAs of uPAR is significantly down-regulated . Targeted down-regulation of uPAR inhibits the malignant biological behaviors including proliferation and migration of colorectal cancer cells.
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