靶向下调uPAR抑制大肠癌细胞恶性生物学行为

Targeted down-regulation of uPAR inhibits the malignant biological behaviors of colorectal cancer cells

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 targeteddown-regulation the expression of uPAR by the specific uPAR-siRNA. Methods： Three pairs of specific uPAR-siRNA wereconstructed and transfected into LOVO colorectal cancer cell line. The interference efficiencies of uPAR-siRNAs weredetected by Western Blot. The uPAR-siRNA with the highest interference efficiency was selected for the followingexperiments. The proliferation of colorectal cancer cells after transfection with uPAR-siRNA was detected by flow cytometryand CCK-8 assay. The migration of coloreetal cancer cells after transfection with uPAR-siRNA was determinated byTranswell chamber system and Wound healing assay. Results： The expressions of uPAR in the colorectal cancer cells weresignificantly down-regulated after transfection with uPAR-siRNAs, and uPAR-siRNA-3 caused the most reductinn of uPAR.After transfeetion with uPAR-siRNA-3 into colorectal cancer cells, the number in Gt phase was significantly increased, butthe 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 issignificantly down-regulated. Targeted down-regulation of uPAR inhibits the malignant biological behaviors includingproliferation and migration of colorectal cancer cells.