过表达FOXN3对食管癌细胞生物学的影响

Effect of overexpression of FOXN3 on the biology of esophageal carcinoma cells

目的探讨过表达叉头框蛋白N3(FOXN3)对食管癌细胞增殖、凋亡、迁移和侵袭的影响。方法通过RT-qPCR实验检测正常食管上皮细胞HEEC、食管癌细胞Eca-109、KYSE-30、TE-1中FOXN3的表达水平,选取表达量最低的Eca-109细胞用于后续实验,通过Lipofectamine2000将pcDNA3.1-FOXN3、pcDNA3.1转染食管癌细胞Eca-109,作为FOXN3过表达组和空载体对照组,对照组细胞不做处理。通过CCK8检测各组细胞增殖情况,流式细胞仪检测各组细胞凋亡及细胞周期,Transwell检测各组细胞侵袭水平,细胞划痕实验检测各组细胞迁移水平,Western blot检测各组细胞cleaved-Caspase-3、Ki-67、MMP-2、Wnt1和β-catenin蛋白表达水平。结果FOXN3在食管癌细胞中低表达(P<0.05),过表达FOXN3使食管癌细胞增殖、迁移和侵袭能力减弱,相关蛋白Ki-67、MMP-2表达显著下降(P<0.05),凋亡率及凋亡相关蛋白cleaved-Caspase-3显著升高(P<0.05),细胞周期受到阻滞,Wnt1和β-catenin蛋白相对表达量显著下降(P<0.05)。结论过表达FOXN3能够抑制食管癌细胞增殖、迁移和侵袭,并促细胞凋亡,其机制可能与过表达FOXN3抑制Wnt信号通路有关。

Objective To investigate the effects of overexpression of Fork head Box N3(FOXN3)on the proliferation,apoptosis,migration and invasion of esophageal carcinoma cells.Methods The expression levels of FOXN3 in normal esophageal epithelial cells HEEC,esophageal cancer cells Eca-109,KYSE-30,and TE-1 was detected by RT-qPCR,and the Eca-109 cells with the lowest expression level were selected for subsequent experiments.Lipofectamine2000 transfected pcDNA3.1-FOXN3 and pcDNA3.1 into esophageal carcinoma cells Eca-109 as FOXN3 overexpression group and empty vector control group.The control group cells were not treated.Cell proliferation in each group was detected by CCK8,apoptosis and cell cycle were detected by flow cytometry,cell invasion level was detected by Transwell,cell migration level was detected by cell scratching experiment,and protein expression levels of cleaved-Caspase-3,Ki-67,MMP-2,Wnt1 andβ-catenin were detected by Western-blot.Results FOXN3 expressed lower in esophageal cancer cells(P<0.05).FOXN3 overexpression reduces the proliferation,migration,and invasion of esophageal cancer cells,significantly decreased Ki-67,MMP-2 protein expression(P<0.05).The apoptosis rate and apoptosis related proteins cleaved-Caspase-3 significantly increased(P<0.05).The blockage of cell cycle,Wnt1 andβ-catenin protein relative expression was significantly decreased(P<0.05).Conclusion FOXN3 overexpression can inhibit the proliferation,migration and invasion of esophageal carcinoma cells and promote apoptosis,which may be related to the inhibition of Wnt signaling pathway by FOXN3 overexpression.