AKT抑制剂对结肠癌细胞生物活性及β-catenin信号的影响  

Effects of AKT inhibitors on the physiological activity of colon cancer cells and β-catenin signal

作　　者：杜光红[1] 余雪莲[1] 陈韵[1] 张彤彤[2] 蔡玉婷 DU Guang-Hong;YU Xue-Lian;CHEN Yun(Sichuan Academy of Medical Sciences.Sichuan Provincial People's Hospital,Chengdu 610072,Sichuan,China)

机构地区：[1]四川省医学科学院四川省人民医院,四川成都610072 [2]重庆医科大学第一附属医院消化内科

出　　处：《中国老年学杂志》2022年第12期2978-2983,共6页Chinese Journal of Gerontology

基　　金：国家自然科学基金(81502075)。

摘　　要：目的 分析丝氨酸/苏氨酸激酶(AKT)抑制剂对结肠癌细胞生物活性及β-catenin信号的影响。方法 取对数生长的5×10^(3)的SW480细胞,分别加入0.0、2.5、5.0、10.0μmol/L的MK2206干预24 h,采用显微镜观察SW480细胞形态;CCK-8检测存活率;克隆实验检测细胞克隆数目;采用Hoechst荧光染色检测凋亡率;分别采用Western印迹及聚合酶链反应(PCR)检测细胞中β-catenin、p-AKT蛋白及mRNA表达。结果 2.5、5.0、10.0μmol/L处理下的SW480细胞均与0.0μmol/L组相比差距较大(P<0.05),5.0μmol/L处理下的SW480存活率低于2.5μmol/L(P<0.05),10.0μmol/L处理下的SW480存活率最低,且具有时间剂量依赖性,2.5μmol/L组SW480细胞克隆数目显著低于0.0μmol/L组(t=3.080,P<0.05),5.0μmol/L组显著低于2.5μmol/L组(t=5.385,P<0.05),10.0μmol/L组显著低于5.0μmol/L组(t=13.96,P<0.05);2.5μmol/L组SW480细胞凋亡率显著高于0.0μmol/L组(t=13.220,P<0.05),5.0μmol/L细胞凋亡率显著高于2.5μmol/L组(t=6.029,P<0.05),10.0μmol/L组SW480细胞凋亡率显著高于5.0μmol/L组(t=8.402,P<0.05),2.5μmol/L组SW480细胞β-catenin和p-AKT蛋白水平显著低于0.0μmol/L组(t_(β-catenin)=3.184,t_(p-AKT)=3.060,均P<0.05),5.0μmol/L组β-catenin和p-AKT蛋白水平显著低于2.5μmol/L组(t_(β-catenin)=3.275,t_(p-AKT)=8.227,均P<0.05),10.0μmol/L组β-catenin和p-AKT蛋白水平显著低于5.0μmol/L组(t_(β-catenin)=5.039,t_(p-AKT)=5.477,P<0.05),2.5μmol/L组SW480细胞β-catenin和AKT mRNA均显著低于0.0μmol/L组(t_(β-catenin)=4.756,t_(p-AKT)=3.132,均P<0.05),5.0μmol/L组β-catenin和AKT mRNA显著低于2.5μmol/L组(t_(β-catenin)=8.227,t_(p-AKT)=3.811,P<0.05),10.0μmol/L组β-catenin和AKT mRNA均显著低于5.0μmol/L组(t_(β-catenin)=9.959,t_(p-AKT)=6.235,P<0.05)。结论 AKT抑制剂MK2206能够减少人结肠癌细胞株SW480生理活动,降低存活率,加快凋亡,能够显著抑制β-catenin、p-AKT活性,随着MK2206浓度增加而降低。Objective To analyze the effects of AKT inhibitors on the physiological activity of colon cancer cells andβ-catenin signal.Methods 5×103 SW480 cells grown logarithmically were taken and added 0.0,2.5,5.0,10.0μmol/L MK2206 to intervene for 24 h,and observe the morphology of SW480 cells with a microscope;CCK-8 test was used to determine survival rate;cloning experiment was used to detect the number of cell clones;Hoechst fluorescence staining was used to detect the apoptosis rate;the protein and mRNA expressions ofβ-catenin and p-AKT were detected by Western blot and PCR,respectively.Results The SW480 cells treated with 2.5,5.0,10.0μmol/L were significantly different from the 0.0μmol/L concentration group(P<0.05),and the survival rate of SW480 cells treated with 5.0μmol/L was lower than that of 2.5μmol/L(P<0.05),the survival rate of SW480 under 10.0μmol/L treatment was the lowest,and it was time-dose-dependent.The number of SW480 cell clones under 2.5μmol/L treatment was less than that of 0.0μmol/L concentration(t=3.080,P<0.05),and the number of 5.0μmol/L cell clones was less than that of 2.5μmol/L(t=5.385,P<0.05).The number of clones in 10.0μmol/L SW480 cells was significantly less than that of 5.0μmol/L group(t=13.96,P<0.05);the apoptosis rate of SW480 cells under 2.5μmol/L treatment was significantly higher than that of 0.0μmol/L concentration(t=13.220,P<0.05),5.0μmol/L cell apoptosis rate was higher than that of 2.5μmol/L group(t=6.029,P<0.05),10.0μmol/L SW480 cell apoptosis rate was significantly higher than that of 5.0μmol/L group(t=8.402,P<0.05).Theβ-catenin and p-AKT protein levels of SW480 cells treated with 2.5μmol/L were significantly lower than those in the 0.0μmol/L concentration(t_(β-catenin)=3.184,t p-AKT=3.060,both P<0.05),theβ-catenin and p-AKT protein levels of SW480 cells treated with 5.0μmol/L were lower than those in the 2.5μmol/L group(t_(β-catenin)=3.275,t p-AKT=8.227,both P<0.05),theβ-catenin and p-AKT protein levels of SW480 cells treated with 10.0μmol/L were

关 键 词：结肠癌 AKT抑制剂 MK2206 细胞形态 存活率 Β连环素 丝氨酸/苏氨酸激酶 

分 类 号：R574.62[医药卫生—消化系统]