激活Wnt信号通路阻滞沉默ZEB2对结直肠癌细胞增殖抑制和凋亡促进作用  

Activation of Wnt signaling pathway inhibits proliferation and apoptosis of colorectal cancer cells by silencing ZEB2

作　　者：杨洲[1] 刚海菊[2] 覃先蓬[1] 贾贵清[1] 王波[1] 杨春[3] YANG Zhou;GANG Hal-ju;OIN Xian- peng(Department of Gastrointestinal Surgery,Sichuan Provincial Peoples Hospital,Chengdu Sichuan 610072,China;Nursing Office,Chengdu Vocational and Technical College,Chengdu Sichuan 610041,China.)

机构地区：[1]四川省医学科学院.四川省人民医院胃肠外科,四川成都610072 [2]成都职业技术学院医护分院护理教研室,四川成都610041 [3]四川省医学科学院..四川省人民医院胃肠外科,四川成都610072

出　　处：《临床和实验医学杂志》2018年第16期1717-1722,共6页Journal of Clinical and Experimental Medicine

基　　金：四川省卫计委科研立项课题项目(编号:17PJ109)

摘　　要：目的研究下调结直肠癌细胞中E盒结合锌指蛋白2(ZEB2)的表达对细胞增殖的影响。方法 ZEB2 shRNA组、ZEB2 shRNA+氯化锂(LiCl)组细胞用感染ZEB2 shRNA慢病毒之后的SW48细胞,shRNA-NC组细胞用感染shRNA-control慢病毒之后的SW48细胞,ZEB2 shRNA+Li Cl组细胞在实验开始时在细胞培养液中添加20 mmol/L的Wnt/β-catenin通路激活剂LiCl。对照组细胞不作任何处理。以实时荧光PCR和Western blot检测结肠细胞中ZEB2的表达水平。用Western blot法测定下调ZEB2后的结直肠癌细胞中Wnt信号通路蛋白β-连环蛋白(β-catenin)、c-Myc表达水平。用Wnt信号通路激活剂处理下调ZEB2后的结直肠癌细胞,CCK8检测细胞增殖,流式细胞术检测细胞凋亡,Western blot检测细胞中凋亡相关蛋白剪切型Caspase-9(c-caspase-9)、剪切型Caspase-3(ccaspase-3)水平。结果 ZEB2在结直肠癌细胞中的表达水平高于正常结肠细胞。ZEB2 shRNA慢病毒感染可以下调结直肠癌细胞中ZEB2的表达。沉默ZEB2后的结直肠癌细胞的增殖能力降低,细胞凋亡增多,细胞中c-caspase-9、c-caspase-3蛋白水平升高,β-catenin、c-Myc蛋白水平降低。Wnt信号通路激活剂可以逆转沉默ZEB2后结直肠癌细胞增殖抑制和凋亡促进作用。结论沉默ZEB2诱导结直肠癌细胞凋亡并抑制细胞增殖,而Wnt信号通路激活剂可以逆转沉默ZEB2的这种作用。Objective To stuudy the effect of down regulation of ZEB2 expression on cell proliferation in colorectal cancer cells. Methods ZEB2 shRNA group,ZEB2 shRNA ＋ LiCl group cells were infected with ZEB2 shRNA lentivirus SW48 cells,shRNA-NC group cells were infected with shRNA-control lentivirus SW48 cells,and ZEB2 shRNA ＋ LiCl group cells were at the start of the experiment in cells 20 mmol/L of Wnt/β-catenin pathway activator LiCl was added to the culture medium. Control group cells did not do anything. The expression levels of ZEB2 in normal colonic cells and colorectal cancer cells were detected by Realtime PCR and Western blot. Using ZEB2 shRNA lentivirus and shRNA control lentivirus to infect colorectal cancer cells,the expression level of ZEB2 in these cells was detected by the above methods. The expression level of Wnt signal pathway protein-catenin and c-Myc in colorectal cancer cells after down-regulation of ZEB2 was determined by Western blot. Wnt signaling pathway activator was used to treat colorectal cancer cells after downregulation of ZEB2. CCK8 detection of cell proliferation,cell apoptosis was detected by flow cytometry,Western blot was used to detect the levels of apoptosis related proteins c-caspase-9 and c-caspase-3. Results The expression level of ZEB2 in colorectal cancer cells was higher than that in normal colon cells,ZEB2 shRNA lentivirus infection can downregulate the expression of ZEB2 in colorectal cancer cells. After silencing ZEB2,the proliferation ability of colorectal cancer cells decreased,apoptosis increased,the level of c-caspase-9 and c-caspase-3 protein increased in cells,the levels of β-catenin and c-Myc protein were decreased. Wnt signaling pathway activator could reverse the proliferation inhibition and apoptosis promotion of colorectal cancer cells after ZEB2 silencing. Conclusion The activation of Wnt signaling pathway blocked the inhibition of ZEB2 on the proliferation and apoptosis of colorectal cancer cells.

关 键 词：结直肠癌 ZEB2 增殖 WNT信号通路 

分 类 号：R735.34[医药卫生—肿瘤]