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目的:探讨人成纤维细胞生长因子3(Fibroblast growth factor3,FGFR3)基因沉默对人类肺腺癌A549细胞侵袭能力及其对基质金属蛋白酶9(Matrix metaloproteinases 9,MMP9)基因表达的影响。方法:细胞分为3组:A组:实验组,即FGFR3特异性小干扰RNA(Small interfering RNA,siRNA)(siRNA-FGFR3)干扰组;B组:阴性对照组,即FGFR3非特异性阴性对照siRNA(siRNA-NC)干扰组;C组:空白对照组,无siRNA干扰;通过核酸转染试剂脂质体Lipofectamine TM2000(Lipo2000)转染A549细胞;倒置荧光显微镜观察Lipo2000转染效率;转染后A549细胞的侵袭能力用Transwell实验检测;实时荧光定量聚合酶链反应(Real-time quantitative polymerase chain reaction,Real-time PCR)用于检测转染前后FGFR3及MMP9 m RNA的表达水平。结果:Lipo2000介导的FAM-siRNA对肺腺癌A549细胞的转染效率可达80%;在转染36 h后,Transwell实验结果显示A组较B组、C组侵袭能力显著降低(P<0.01)。Real-time PCR结果显示,A组较B、C组的FGFR3和MMP9基因表达量明显下调(P<0.01)。结论:FGFR3基因沉默可明显抑制肺腺癌A549细胞的侵袭能力,并能下调MMP9表达。为肺癌的治疗提供了新的靶点。
Objective: To investigate the effect of FGFR3 gene silencing on invasiveness of human lung adenocarcinoma A549 cells and its effect on matrix metalloproteinase 9 (MMP9) gene expression. Methods: The cells were divided into 3 groups: group A: experimental group, which was interference group of FGFR3-specific small interfering RNA (siRNA) (siRNA-FGFR3); group B: negative control group of FGFR3 non-specific negative control siRNA (siRNA-NC) interference group; group C: blank control group, no siRNA interference; transfection of A549 cells by lipofectamine TM2000 (Lipo2000) by nucleic acid transfection reagent; transfection efficiency of Lipo2000 by inverted fluorescence microscope; The invasion ability of cells was detected by Transwell assay. Real-time quantitative polymerase chain reaction (Real-time PCR) was used to detect the expression of FGFR3 and MMP9 mRNA before and after transfection. Results: The transfection efficiency of Lipo2000-mediated FAM-siRNA on lung adenocarcinoma A549 cells was up to 80%. Transwell assay showed that the transfection efficiency of group A was significantly lower than that of group B and C (P < 0.01). Real-time PCR results showed that the expression of FGFR3 and MMP9 in group A was significantly lower than that in group B and C (P <0.01). Conclusion: Silencing of FGFR3 gene significantly inhibits the invasiveness of lung adenocarcinoma A549 cells and down-regulates the expression of MMP9. It provides a new target for the treatment of lung cancer.