【摘要】 目的利用针对Lipocalin-2的小干扰RNA(siRNA),观察Lipocalin-2基因沉默对人肺癌A549细胞MMP-9表达及侵袭、迁移能力的影响。方法采用Real-time PCR和Western blot检测Lipocalin-2在3种肺癌细胞株(A549、NCI-H661、NCI-H446)及正常人支气管上皮细胞(HBE)的表达情况。设计并构建靶向Lipocalin-2的siRNA转染A549细胞,采用Real-time PCR和Western blot检测转染效率及Lipocalin-2基因沉默对MMP-9 mRNA和蛋白表达的影响。Transwell小室检测细胞侵袭能力。细胞划痕实验检测细胞迁移能力。结果 Lipocalin-2 mRNA和蛋白在肺癌A549、NCI-H661、NCI-H446细胞株中的表达水平均显著高于HBE细胞,差异有统计学意义(P<0.05)。siRNA-Lipocalin-2转染组Lipocalin-2 mRNA和蛋白表达水平与阴性对照组和空载体对照组比较,差异有统计学意义(P<0.05);siRNA-Lipocalin-2转染组MMP-9 mRNA和蛋白表达水平与阴性对照组和空载体对照组比较,差异有统计学意义(P<0.05);siRNA-Lipocalin-2转染组穿膜细胞数、平均迁移距离与阴性对照组和空载体对照组比较,差异有统计学意义(P<0.05)。结论 Lipocalin-2基因沉默通过下调MMP-9表达降低了人肺癌A549细胞的侵袭和迁移能力。更多还原

【Abstract】 Objective To investigate the effects of gene silencing of Lipocalin-2 on the expression of MMP-9 and metastasis and invasion abilities of human lung cancer A549 cells in vitro.Methods The expression levels of Lipocalin-2 mRNA and protein in 3 kinds of human lung cancer cell lines( A549,NCI-H661,NCI-H446) and healthy human bronchial epithelial cells( HBE) were detected by Real-time PCR and Western Blot,respectively.The targeting Lipocalin-2 siRNA was established and transfected into A549 cells via lentivirus.Transfection efficiency and the expression levels of MMP-9 mRNA and protein were detected by Real-time PCR and Western Blot.Moreover the invasive ability and migration ability of A549 cells were detected by Transwell assay and scratch test,respectively.Results The expression levels of Lipocalin-2 mRNA and protein in A549,NCI-H661,NCI-H446 cells were significantly higher than those in HBE cells(P<0.05),furthermore,there were significant differences in the expression levels of Lipocalin-2 mRNA and protein between siRNA-Lipocalin-2transfection group and negative control group as well as empty vector control group(P<0.05).Besides there were significant differences in the cell number with membrane tresis and average migration distance between siRNA-Lipocalin-2 transfection group and negative control group as well as empty vector control group(P<0.05).Conclusion Lipocalin-2 gene silencing can decrease metastasis and invasion abilities of human lung cancer A549 cells in vitro through down-regulating the expression levels of MMP-9.更多还原