ZEB2调控胰腺癌PANC-1细胞迁移和侵袭的实验研究  

ZEB2 Regulates the Migration and Invasion of PANC-1 Pancreatic Cancer Cells:An Experimental Study

作　　者：张荣花 黄金平 李景武[2] 李亚琦 崔笑妍 熊亚南 刘艳坤 章广玲 ZHANG Rong-hua;HUANG Jin-ping;LI Jing-wu;LI Ya-qi;CUI Xiao-yan;XIONG Ya-nan;LIU Yan-kun;ZHANG Guangling(Hebei Provincial Key Laboratory for Chronic Diseases,School of Basic Medical Sciences,North China University of Science and Technology,Tangshan 063210,China;Hebei Provincial Key Laboratory of Molecular Oncology,Cancer Research Institute of Tangshan People's Hospital,Tangshan 063001,China;Tangshan City Key Laboratory of Precision Medical Testing,Tangshan 063001,China;Hebei Provincial Key Laboratory of Medical-Industrial Integration Precision Medicine,School of Clinical Medicine,North China University of Science and Technology,Tangshan 063000,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]唐山市人民医院肿瘤研究所河北省分子肿瘤学重点实验室,唐山063001 [3]唐山市精准医学医学检测重点实验室,唐山063001 [4]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：《四川大学学报（医学版）》2023年第3期558-564,共7页Journal of Sichuan University(Medical Sciences)

基　　金：河北省自然科学基金资助项目(No.H2021209026、No.H2021105019);河北省人力资源和社会保障厅项目(No.C20210340);河北省重点研发计划项目(No.213777115D);河北省财政厅项目(冀财预复[2020]397号)资助。

摘　　要：目的研究锌指E-box结合同源异型盒2基因(zinc finger E-box binding homeobox transcription factor-2,ZEB2)对胰腺癌PANC-1细胞增殖、集落形成、迁移和侵袭能力及上皮-间质转化(epithelial-mesenchymal transition,EMT)过程的影响。方法分析癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中胰腺癌组织和癌旁组织中ZEB2的表达。将胰腺癌PANC-1细胞分为si-NC组、si-ZEB2组、pcDNA3.1组和pcDNA3.1-ZEB2组,采用qRT-PCR技术和Western blot法验证ZEB2敲降或过表达的有效性。CCK-8实验、集落形成实验、划痕实验和Transwell实验分别检测ZEB2对PANC1细胞的增殖、集落形成、迁移和侵袭的影响。利用qRT-PCR技术和免疫荧光实验检测细胞中EMT标志物E-cadherin、vimentin的表达水平。通过STRING网站预测与ZEB2具有相互作用的蛋白。结果TCGA数据库分析显示,与癌旁组织相比,胰腺癌组织中ZEB2表达水平明显升高(P<0.05)。与si-NC组相比,si-ZEB2组PANC-1细胞的增殖、集落形成、迁移、侵袭能力减弱;与pcDNA3.1组相比,pcDNA3.1-ZEB2组PANC-1细胞的增殖、集落形成、迁移、侵袭能力增强(均P<0.05)。qRT-PCR技术和免疫荧光实验结果提示,与si-NC组相比,si-ZEB2组PANC-1细胞的上皮标志物E-cadherin mRNA表达升高,而间质标志物vimentin mRNA和蛋白表达降低;与pcDNA3.1组相比,pcDNA3.1-ZEB2组PANC-1细胞中上皮标志物Ecadherin mRNA表达减少,间质标志物vimentin mRNA和蛋白表达增加(均P<0.05)。STRING网站预测出10个蛋白与ZEB2作用密切。结论过表达ZEB2能够促进胰腺癌PANC-1细胞的迁移、侵袭和EMT进程。Objective To investigate the effects and mechanisms of zinc finger E-box binding homeobox transcription factor-2(ZEB2)on the proliferation,colony formation,migration,and invasion abilities and the epithelialmesenchymal transition(EMT)of PANC-1 cells,a human pancreatic cancer cell line.Methods Data on the expression of ZEB2 in pancreatic cancer tissues and paracancerous tissues from The Cancer Genome Atlas(TCGA)database were analyzed.PANC-1 pancreatic cancer cells were divided into si-NC group,si-ZEB2 group,pcDNA3.1 group,and pcDNA3.1-ZEB2 group.qRT-PCR and Western blot were conducted to confirm the effectiveness of ZEB2 knockdown or overexpression.CCK-8,colony formation,wound healing,and Transwell assays were conducted to examine the effects of ZEB2 on the proliferation,colony formation,migration,and invasion of PANC-1 cells.qRT-PCR and immunofluorescence assays were performed to examine the expression of E-cadherin and vimentin,the EMT markers,in the cells.Prediction of proteins interacting with ZEB2 was made through the STRING database.Results TCGA database analysis showed that the expression level of ZEB2 in pancreatic cancer tissues was significantly higher than that in adjacent tissues(P<0.05).Compared with those of cells in the control group,the proliferation,colony formation,migration,and invasion of cells in the si-ZEB2 group were decreased(P<0.05).Compared with those of cells in the pcDNA3.1 group,the proliferation,colony formation,migration and invasion of cells in the pcDNA3.1-ZEB2 group were increased(all P<0.05).According to the results of qRT-PCR and immunofluorescence assays,compared with those of the si-NC group,the expression of Ecadherin mRNA,an epithelial marker,in the si-ZEB2 group increased,while the expression of vimentin mRNA,an mesenchymal marker,and the protein decreased.Compared with those of the pcDNA3.1 group,the expression of Ecadherin mRNA in the PANC-1 cells of the pcDNA3.1-ZEB2 group decreased,while the expression of vimentin mRNA and the protein increased(all P<0.05).Analysis with

关 键 词：ZEB2 胰腺癌 PANC-1细胞 增殖 迁移 侵袭 

分 类 号：R735.9[医药卫生—肿瘤]