FOXO3抑制膀胱癌细胞生长的体外实验研究  

Inhibitory effect of FOXO3 on the growth of bladder cancer cells in vitro

作　　者：何笑凯[1] 康郑军[1] 柴蕴珠[2] HE Xiaokai;KANG Zhengjun;CHAI Yunzhu(Department of Urologic Surgery,the Fifth Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,He’nan,China;Department of Gastroenterology,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,He’nan,China)

机构地区：[1]郑州大学第五附属医院泌尿外科,郑州450052 [2]郑州大学第一附属医院消化内科,郑州4500000

出　　处：《癌症进展》2019年第7期784-787,共4页Oncology Progress

摘　　要：目的研究叉头框转录因子O3(FOXO3)对膀胱癌细胞生长的影响及机制。方法将人膀胱癌T24细胞转染FOXO3-pcDNA3.1和pcDNA3.1,分别记为FOXO3组、Vector组,将没有转染的T24细胞记为对照组。反转录-聚合酶链反应(RT-PCR)检测FOXO3 mRNA表达情况,蛋白质印迹(Western blot)法检测FOXO3蛋白表达情况,噻唑蓝(MTT)法检测细胞增殖能力,细胞克隆形成实验检测细胞克隆形成能力,Western blot法检测β-链蛋白(β-catenin)、蛋白激酶B(AKT)、磷酸化蛋白激酶B(p-AKT)蛋白表达情况。结果 Vector组细胞中FOXO3mRNA和蛋白相对表达量与对照组比较,差异均无统计学意义(P﹥0.05);FOXO3组细胞中FOXO3 mRNA和蛋白相对表达量均明显高于对照组,差异均有统计学意义(P﹤0.01)。Vector组细胞光密度(OD)值、细胞克隆数目与对照组比较,差异均无统计学意义(P﹥0.05);FOXO3组细胞OD值、细胞克隆数目均低于对照组,差异均有统计学意义(P﹤0.05)。Vector组细胞中β-catenin、p-AKT、AKT蛋白相对表达量与对照组比较,差异均无统计学意义(P﹥0.05);FOXO3组细胞中AKT蛋白相对表达量与对照组比较,差异无统计学意义(P﹥0.05);FOXO3组细胞中β-catenin、p-AKT蛋白相对表达量均低于对照组,差异均有统计学意义(P﹤0.05)。结论 FOXO3能够在体外抑制膀胱癌细胞生长,下调β-catenin和p-AKT蛋白表达。Objective To investigate the effect and mechanism of forkhead box O3 (FOXO3) on the growth of bladder cancer cells.Method Human bladder cancer T24 cells were transfected with FOXO3-pcDNA3.1 and pcDNA3.1,as FOXO3 group and Vector group,respectively,besides,the non-transfected cells were included as control group.Reverse transcription- polymerase chain reaction (RT- PCR) was utilized to detect the expression of FOXO3 mRNA,and the FOXO3 protein level were determined by Western blot.Cell proliferation was detected by methyl thiazolyldiphenyl tetrazolium (MTT) assay,and the cell cloning assay was applied to evaluate the clonogenic ability of cells,the expression of β-catenin,protein kinase B (AKT) and p-AKT protein were assessed using Western blot.Result The relative expression level of FOXO3 mRNA and protein were similar between Vector group and control group (P>0.05);while those in FOXO3 group were significantly higher compared with control group,and the difference were of statistical significance (P<0.01).The optical density (OD) value,and number of cell clone formation were comparable between Vector group and control group (P>0.05);while those in FOXO3 group were evidently lower compared with control group (P<0.05). As to the relative expression level of β-catenin,p-AKT and AKT proteins,similar level were observed in Vector group and control group (P>0.05),while in FOXO3 group,the relative expression level of AKT protein was similar with that in control group (P>0.05),though β-catenin and p-AKT proteins were lower compared with control group (P<0.05).Conclusion FOXO3 can inhibit the growth of bladder cancer cells in vitro,and lower the expression of β-catenin and p-AKT proteins.

关 键 词：膀胱癌 FOXO3 克隆 增殖 

分 类 号：R737.14[医药卫生—肿瘤]