【摘要】 目的探讨头框转录因子O家族4(class O of forkhead box transcriptionfactor 4,FOXO4)对喉癌细胞增殖凋亡能力的影响。方法应用Western blot法检测喉癌组织及对应癌旁组织中FOXO4的表达水平。细胞转染FOXO4过表达载体(p-EGFP-C1/FOXO4组)和空载体(p-EGFP-C1组),同时设置未转染组,未转染组中只加入转染试剂。Western blot法检测转染后细胞中FOXO4蛋白水平。四甲基偶氮唑蓝(MTT)检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中活化的含半胱氨酸的天冬氨酸蛋白水解酶3(Cleaved Caspase-3)、Caspase-3、活化的含半胱氨酸的天冬氨酸蛋白水解酶9(Cleaved Caspase-9)、Caspase-9、β-连环蛋白(β-catenin)、Wnt1表达水平。用Wnt/β-catenin信号通路激活剂作用于转染p-EGFP-C1/FOXO4后的喉癌细胞(激活剂组),检测细胞增殖、凋亡情况。结果 FOXO4在喉癌组织中表达水平明显低于癌旁组织(P=0.000)。p-EGFP-C1/FOXO4组细胞中FOXO4表达水平明显高于未转染组(P=0.000)。p-EGFP-C1/FOXO4组细胞存活率及β-catenin、Wnt1水平明显低于未转染组(P=0.002,P=0.004,P=0.006),细胞凋亡率及Cleaved Caspase-3、Caspase-3、Cleaved Caspase-9、C a s p a s e-9表达水平均明显高于未转染组(P=0.0 0 2,P=0.001,h P<0.05,P=0.004,j P<0.05)。Wnt/β-catenin信号通路激活剂可以部分逆转FOXO4抑增殖和促凋亡作用。结论 FOXO4能够促进人喉癌细胞凋亡,抑制喉癌细胞增殖,作用机制可能与Wnt/β-catenin信号通路有关。更多还原

【Abstract】 OBJECTIVE To investigate the effect of FOXO4 on proliferation and apoptosis of laryngeal carcinoma cells.METHODS The expression of FOXO4 in laryngeal carcinoma tissues and adjacent tissues was detected by Western blot.Laryngeal carcinoma cells were transfected with FOXO4 overexpression vector(p-EGFP-C1/FOXO4 group) and empty vector(p-EGFP-C1 group),while the non transfection group was established,and transfection reagent was only added in the non transfection group.The level of FOXO4 protein in transfected cells was detected by Western blot.Cell proliferation was detected by MTT,apoptosis was detected by flow cytometry.The levels of Cleaved Caspase-3,Caspase-3,Cleaved Caspase-9,β-catenin,Wnt1 were detected by Western blot.The transfected laryngeal carcinoma cells with p-EGFP-C1/FOXO4 was activated by Wnt/β-catenin signal pathway activator(activator group),and the proliferation and apoptosis of the cells were detected.RESULTS The expression level of FOXO4 in laryngeal carcinoma tissues was significantly lower than that in adjacent tissues(P=0.000).The expression level of FOXO4 in p-EGFP-C1/FOXO4 group was significantly higher than that in non transfection group(P = 0.000).The sur vival rate and the expression levels of β-catenin and Wnt1 in p-EGFP-C1/FOXO4 group were significantly lower than those in the non transfection group(P =0.002,P =0.004,P =0.006).Cell apoptosis rate and expression levels of Cleaved Caspase-3,Caspase-3,Cleaved Caspase-9,Caspase-9 in p-EGFP-C1/FOXO4 group were significantly higher than those in non transfection group(P =0.002,P =0.001,h P <0.05,P =0.004,j P <0.05).Wnt/β-catenin signaling pathway activator can partly reverse the proliferation inhibition and apoptosis promoting effect of FOXO4.CONCLUSION FOXO4 could promote t he apoptosis of human laryngeal carcinoma cells and inhibit the proliferation of laryngeal cancer cells.The mechanism may be related to the signal pathway of Wnt/β-catenin.更多还原