CXXC4调控胰腺癌PANC-1细胞迁移和侵袭的实验研究  

Experimental study on the regulation of CXXC4 on the migration and invasion of PANC-1 pancreatic carcinoma cells

作　　者：王梅梅 崔笑妍 张亚楠 周静 张荣花 熊亚南 刘志勇 章广玲 WANG Meimei;CUI Xiaoyan;ZHANG Yanan;ZHOU Jing;ZHANG Ronghua;XIONG Yanan;LIU Zhiyong;ZHANG Guangling(University of Science and Technology,Tangshan 063210,Hebei,China;North China University of Science and Technology Affiliated Hospital and Health Science Center,Tangshan 063000,Hebei,China;Hebei Provincial Key Laboratory of Medical-Industrial Integration Precision Medicine,North China University of Science and Technology Affiliated Hospital,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]华北理工大学附属医院医学部,唐山063000 [3]华北理工大学附属医院,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》2023年第12期1233-1241,共9页Journal of Medical Research & Combat Trauma Care

基　　金：河北省自然科学基金(H2021209026,H2023209047)。

摘　　要：目的探究CXXC指蛋白4(CXXC4)对胰腺癌PANC-1细胞迁移、侵袭及上皮-间充质转化(EMT)进程的影响。方法组织芯片免疫组织化学染色实验检测14份癌旁胰腺组织和87份胰腺癌组织中CXXC4表达情况。采用免疫荧光实验检测人正常胰腺导管上皮细胞HPNE及人胰腺癌细胞PANC-1、AsPC-1和BxPC-3中CXXC4蛋白的表达水平。采用Western blot实验检测CXXC4敲降和过表达的转染有效性,划痕和Transwell实验分析敲降或过表达CXXC4对PANC-1细胞迁移、侵袭的影响。采用Western blot和免疫荧光实验检测细胞EMT标志物E-cadherin、Vimentin和ZEB2的表达水平。通过STRING网站预测与CXCC4有相互作用的蛋白,筛选出的蛋白进行GO和KEGG富集分析。结果免疫组化分析结果表明,CXXC4在胰腺癌组织中的表达水平低于癌旁胰腺组织(P<0.05)。免疫荧光实验结果显示,与HPNE细胞相比,CXXC4在PANC-1、AsPC-1和BxPC-3中的荧光强度均显著降低(均P<0.05)。划痕和Transwell实验结果显示,与si-NC组相比,si-CXXC4组PANC-1细胞迁移和侵袭能力增强(P<0.05);与pcDNA3.1组相比,pcDNA3.1-CXXC4组PANC-1细胞迁移和侵袭能力减弱(P<0.05)。Western blot和免疫荧光实验结果显示,与si-NC组(结果标准化为1)相比,si-CXXC4组PANC-1细胞上皮样细胞标志物E-cadherin蛋白表达(0.48±0.10)降低(P<0.05),而间充质样标志物Vimentin(1.42±0.06)和ZEB2(1.86±0.06)的表达升高(P<0.05);与pcDNA3.1组(结果标准化为1)相比,pcDNA3.1-CXXC4组PANC-1细胞上皮样细胞标志物E-cadherin蛋白表达(2.21±0.42)升高(P<0.05),而间充质样标志物Vimentin(0.54±0.05)和ZEB2(0.39±0.02)蛋白表达降低(P<0.05)。STRING网站预测出9个蛋白与CXXC4有密切的作用。结论CXXC4抑制胰腺癌PANC-1细胞迁移、侵袭及EMT进程。Objective Previousstudies show that CXXC Finger Protein 4(CXXC4)participates in the development of pancreatic carcinoma,but the underlying mechanism needs further study.This study aimed to investigate the effect of CXXC4 on the migration,invasion and Epithelial-Mesenchymal Transformation(EMT)of pancreatic cancer PANC-1 cells.Methods Immunohistochemistry(IHC)was used to examine the expression level of CXXC4 in 14 para cancer pancreatic tissues and 87 pancreatic cancer tissues.Immuno-fluorescence assay was used to detect CXXC4 protein expression in human normal pancreatic ductal epithelial cells HPNE and human pancreatic cancer cells PANC-1,AsPC-1 and BxPC-3.Western blot was used to confirm the effectiveness of CXXC4 knockdown or overexpression.Wound healing and Transwell assays were used to evaluate the effects of knockdown or overexpression of CXXC4 on PANC-1 cell migration and invasion.Western blot and immunofluorescence assays were used to detect the expression levels of EMT markers E-cadherin,Vimentin and ZEB2.Potential proteins interacting with CXXC4 were predicted through the STRING database.The selected proteins were enriched by GO and KEGG.Results Immunohistochemical analysis showed that the expression level of CXXC4 in pancreatic cancer tissue was lower than that in pancreatic cancer adjacent tissue(P<0.05).The results of immunofluorescence experiments showed that the expression of CXXC4 protein in PANC-1,AsPC-1 and BxPC-3 cells was lower than that in HPNE cells(all P<0.05).Wound healing and Transwell assays showed that PANC-1 cell migration and invasion ability were enhanced in si-CXXC4 group compared with si-NC group(all P<0.05).Compared with pcDNA3.1 group,PANC-1 cell migration and invasion ability were decreased in PCDNA3.1-CXXC4 group(all P<0.05).The results of Western blot and immunofluorescence assays showed that compared with the si-NC group(Normalized 1),the expression of the epithelial marker E-cadherin protein in PANC-1 cells in the si-CXXC4 group(0.48±0.10)was decreased,while the expression o

关 键 词：CXXC指蛋白4 胰腺癌 PANC-1细胞 迁移 侵袭 

分 类 号：R735.9[医药卫生—肿瘤]