FOXC1基因对肺癌A549细胞株生物学特性影响

EFFECT OF FORKHEAD BOX C1 GENE SILENCING ON THE BIOLOGICAL CHARACTERISTICS OF LUNG CANCER A549 CELL LINE

目的探讨沉默叉头框蛋白C1(FOXC1)基因对肺癌A549细胞株生物学特性的影响。方法培养人肺癌A549细胞,随机分为siRNA-FOXC1组、错义siRNA组和空白对照组,应用实时荧光定量PCR技术检测细胞中FOXC1基因表达,Western blot法检测细胞中FOXC1蛋白表达,CCK-8法检测细胞增殖能力,流式细胞术检测细胞凋亡率,划痕实验检测细胞迁移能力,Transwell法检测细胞侵袭能力。结果siRNA-FOXC1组细胞FOXC1mRNA和蛋白相对表达量均低于错义siRNA组和空白对照组,差异均有显著性(F=2 521.827、37.226,P<0.05);siRNA-FOXC1组48、72和96h时细胞存活率均低于错义siRNA组和空白对照组,差异均有显著性(F=22.857～103.502,P<0.05);siRNA-FOXC1组细胞凋亡率高于错义siRNA组和空白对照组,差异有显著性(F=25.844,P<0.05)。划痕实验显示,siRNA-FOXC1组24h后划痕愈合率低于错义siRNA组和空白对照组,差异有显著性(F=70.260,P<0.05)。Transwell实验显示,siRNA-FOXC1组侵袭细胞数低于错义siRNA组和空白对照组,差异有显著性(F=95.879,P<0.05)。结论特异性沉默人肺癌A549细胞中FOXC1基因表达可减少细胞增殖,促进细胞凋亡,有效抑制细胞迁移及侵袭能力。

Objective To investigate the effect of forkhead box C1 （ FOXC1 ） gene silencing on the biological characteristics of lung cancer A549 cell line. Methods Human lung cancer A549 cells were cultured and randomly divided into siRNA-FOXC1 group, scramble siRNA group, and blank control group. Quantitative real-time PCR and Western blot were used to measure the mRNA and protein expression of FOXC1 in cells; CCK-8 assay was used to measure cell proliferation; flow cytometry was used to measure cell apoptosis rate; wound healing assay was used to evaluate cell migration ability; Transwell assay was used to evaluate cell invasion ability. Results The siRNA-FOXC1 group had significantly lower mRNA and protein expression of FOXC1 than the scramble siRNA group and the blank control group （ F =2 521.827,37.226; P 〈0.05）. The siRNA-FOXC1 group had significantly lower survival rates of cells at 48,72, and 96 h than the scramble siRNA group and the blank control group （ F = 22.857- 103.502, P 〈0.05）. The siRNA-FOXC1 group had a significantly higher apoptosis rate than the scramble siRNA group and the blank control group （ F =25.844, P 〈0.05）. The wound healing assay showed that after 24 h, the siRNA-FOXC1 group had a significantly lower wound healing rate than the scramble siRNA group and the blank control group （ F =70.260, P 〈 0.05 ）. The Transwell assay showed that the siRNA-FOXC1 group had a lower number of invasive cells than the scramble siRNA group and the blank control group （ F =95.879, P 〈0.05）. Conclusion Specific silencing of the FOXC1 gene in human lung cancer A549 cells can reduce cell proliferation, promote cell apoptosis, and effectively inhibit cell migration and invasion abilities.