E盒结合锌指蛋白2对AsPC-1细胞增殖和EMT的影响  

Effect of E-box binding zinc finger protein 2 on AsPC-1 cell proliferation and EMT

作　　者：王璐瑶 周静 张荣花 于笑涵 熊亚南 王梅梅 章广玲 刘志勇 WANG Luyao;ZHOU Jing;ZHANG Ronghua;YU Xiaohan;XIONG Yanan;WANG Meimei;ZHANG Guangling;LIU Zhiyong(Hebei Provincial Key Laboratory for Chronic Diseases,School of Basic Medicine,North China University of Science and Technology,Tangshan 063210,Hebei,China;Hebei Provincial Key Laboratory of Medical-Industrial Integra-tion Precision Medicine,School of Clinical Medicine,North China University of Science and Technology,Tangshan 063000,Hebei,China)

机构地区：[1]华北理工大学基础医学院,河北省慢性疾病重点实验室,唐山063210 [2]华北理工大学临床医学院,河北省医工融合精准医疗重点实验室,唐山063000

出　　处：《医学研究与战创伤救治》2024年第8期798-805,共8页Journal of Medical Research & Combat Trauma Care

基　　金：河北省中央引导地方科技发展资金项目(246Z7720G)。

摘　　要：目的探讨E盒结合锌指蛋白2(ZEB2)对AsPC-1胰腺癌细胞增殖、迁移、侵袭和上皮间充质转化(EMT)的影响。方法按照胰腺癌细胞中转染物不同,将细胞分为4组,分别是si-NC、si-ZEB2、pcDNA3.1、pcDNA3.1-ZEB2。TCGA数据库分析ZEB2在胰腺癌组织与癌旁胰腺组织中的表达差异及其与胰腺癌患者预后的关系,并利用Westernblot检测ZEB2在AsPC-1胰腺癌细胞和正常胰腺上皮细胞HPNE中的表达差异。采用qRT-PCR、Westernblot检测ZEB2小干扰RNA(si-ZEB2)及过表达质粒(pcDNA3.1-ZEB2)的转染效率。CCK-8、克隆形成、划痕和Transwell实验检测ZEB2对AsPC-1细胞增殖、克隆形成、迁移和侵袭的影响。qRT-PCR和免疫荧光实验检测ZEB2对AsPC-1细胞中EMT标志物表达的影响。利用生物信息学网站预测与ZEB2有靶定作用的miRNAs并构建ZEB2的蛋白-蛋白互作网络(PPI)。结果ZEB2在胰腺癌组织和AsPC-1胰腺癌细胞中高表达(P<0.05),且其高表达与胰腺癌患者的较差预后相关。qRT-PCR、Westernblot结果表明si-ZEB2和pcDNA3.1-ZEB2能够有效的转染到AsPC-1胰腺癌细胞中(P<0.05)。与si-NC组相比,si-ZEB2组中AsPC-1细胞的增殖、克隆形成、迁移和侵袭能力受到抑制,EMT上皮标志物E-cadherin表达增加、间质标志物Vimentin表达减少(P<0.05)。pcDNA3.1-ZEB2组中AsPC-1细胞的增殖、克隆形成、迁移、侵袭和EMT标志物得到了与上述相反的结果(P<0.05)。共预测到与ZEB2有靶定作用的miRNAs 77个,与ZEB2作用密切的蛋白有10个。结论ZEB2在胰腺癌组织和细胞中高表达且其高表达与胰腺癌患者的较差预后相关,ZEB2作为一种促癌因子,能够促进AsPC-1胰腺癌细胞的增殖、克隆形成、迁移、侵袭和EMT进程。Objective To investigate the effects of E-box binding zinc finger protein 2(ZEB2)on the proliferation,migration,invasion and epithelial-mesenchymal transformation(EMT)of AsPC-1 pancreatic cancer cells.Methods According to different transfection in pancreatic cancer cells,the cells were divided into four groups,namely si-NC,si-ZEB2,pcDNA3.1,PCDNA3.1-ZEB2.The TCGA database was used to analyze the difference of ZEB2 expression in pancreatic cancer tissues and para-cancerous pancreatic tissues and its relationship with the prognosis of pancreatic cancer pa‐tients.Western blot was used to detect the difference of ZEB2 expres‐sion in AsPC-1 pancreatic cancer cells and HPNE in normal pancreat‐ic epithelial cells.The transfection efficiency of ZEB2 small interfer‐ing RNA(si-ZEB2)and overexpressed plasmid(pcDNA3.1-ZEB2)was detected by qRT-PCR and Western blot.The effects of ZEB2 on proliferation,colony formation,migration and invasion of AsPC-1 cells were detected by CCK-8,colony formation,wound healing and Transwell assays.The effect of ZEB2 on the expression of EMT mark‐ers in AsPC-1 cells was detected by qRT-PCR and immunofluorescence assays.Bioinformatics websites were used to predict the miR‐NAs that target ZEB2 and construct the protein-protein interaction network(PPI)of ZEB2.Results ZEB2 was highly expressed in pancreatic cancer tissues and AsPC-1 pancreatic cancer cells(P<0.05),and its high expression was associated with poor prognosis of pancreatic cancer patients.The results of qRT-PCR and Western blot showed that si-ZEB2 and pcDNA3.1-ZEB2 could effectively transfected into AsPC-1 pancreatic cancer cells(P<0.05).Compared with the si-NC group,the proliferation,colony formation,migra‐tion and invasion abilities of AsPC-1 cells in si-ZEB2 group were inhibited,the expression of EMT epithelial marker E-cadherin was in‐creased,and the expression of interstitial marker Vimentin was decreased(P<0.05).In pcDNA3.1-ZEB2 group,the resuts of prolifera‐tion,colony formation,migration,invasion and EMT markers

关 键 词：E盒结合锌指蛋白2 胰腺癌 AsPC-1细胞 增殖 上皮间充质转化 

分 类 号：R735.9[医药卫生—肿瘤]