靶向抑制ZNF281基因表达对直肠癌细胞生长抑制及化疗增敏作用的机制探讨

The mechanism of targeted inhibition of ZNF281 gene expression on growth and chemosensitization of rectal cancer

目的:探讨抑制ZNF281基因表达对直肠癌细胞活力、凋亡率及5-氟尿嘧啶(5-FU)化疗敏感性的影响。方法:以Lipofectamine^(TM) 2000(Lipo2000)为载体,将siRNA转染人直肠癌Colo320细胞,Colo320细胞分为NC组(阴性对照siRNA)、si-ZNF281组(靶向抑制ZNF281表达的小干扰RNA)和si-ZNF281+5-FU组,MTT检测细胞活力,流式细胞仪检测细胞凋亡率,Western blotting检测β-catenin、PCNA和Survivin的蛋白表达。结果:ZNF281 siRNA转染Colo320细胞后,ZNF281表达显著低于空白组(P<0.05)。与NC组比较,si-ZNF281组细胞活力降低,凋亡率升高,β-catenin、PCNA和Survivin的蛋白表达降低,差异均具有统计学意义(P<0.05)。与si-ZNF281组比较,si-ZNF281+5-FU组细胞活力降低,凋亡率升高,β-catenin、PCNA和Survivin的蛋白表达降低,差异均具有统计学意义(P<0.05)。结论:抑制ZNF281基因表达可降低直肠癌细胞活力及诱导细胞凋亡,且可增加5-FU化疗敏感性,机制与抑制Wnt/β-catenin信号通路有关。

Objective:To investigate the effect of inhibiting of ZNF281 gene expression on cell viability,apoptosis rate and 5-fluorouracil chemosensitivity in rectal cancer cells.Methods:Lipofectamine^TM 2000(Lipo2000) was used as the carrier,and siRNA was transfected into Colo320 cells of human rectal cancer.Colo320 cells were divided into NC group(negative control siRNA),si-ZNF281 group(small interference RNA for targeting inhibition of ZNF281 expression) and si-ZNF281+5-FU group.The cell viability was detected by MTT methods.The apoptosis rate was detected by flow cytometry.The protein expression of β-catenin,PCNA and Survivin was detected by Western blotting.Results:The expression of ZNF281 was significantly lower than that the blank group after ZNF281 siRNA was transfected to Colo320 cells(P<0.05).Compared with the NC group,the cell vitality was decreased in si-ZNF281 group,and the apoptosis rate was increased,and the protein expressions of β-catenin,PCNA and Survivin were decreased,and the difference was statistically significant(P<0.05).Compared with the si-ZNF281 group,the cell vitality was decreased in si-ZNF281+5-FU group,and the apoptosis rate was increased,and the protein expressions of β-catenin,PCNA and Survivin were decreased,and the difference was statistically significant(P<0.05).Conclusion:Inhibition of ZNF281 gene expression can inhibit the activity of rectal cancer cells and induce cell apoptosis,and can increase the sensitivity of 5-FU chemotherapy.The mechanism is related to the inhibition of Wnt/β-catenin signaling pathway.