胰腺癌细胞中CXXC4表达水平变化对其增殖、凋亡的影响  

Effect of CXXC4 expression level change in pancreatic cancer cells on proliferation and apoptosis

作　　者：周静 王璐瑶 崔笑妍 张亚楠 张荣花 王梅梅 熊亚南 刘志勇 章广玲 ZHOU Jing;WANG Luyao;CUI Xiaoyan;ZHANG Yanan;ZHANG Ronghua;WANG Meimei;XIONG Yanan;LIU Zhiyong;ZHANG Guangling(Hebei Provincial Key Laboratory for Chronic Diseases,School of Basic Medical Sciences,North China University of Science and Technology,Tangshan,Hebei 063210,China;Affiliated Hospital of North China University of Science and Technology,Tangshan,Hebei 063000,China;Department of Medicine,North China University of Science and Technology,Tangshan,Hebei 063210,China;Hebei Provincial Key Laboratory of Precision Medicine for Medical and Industrial Integration,Clinical School of Medicine,North China University of Science and Technology,Tangshan,Hebei 063015,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,河北唐山063210 [2]华北理工大学附属医院,河北唐山063000 [3]华北理工大学医学部,河北唐山063210 [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,河北唐山063015

出　　处：《重庆医学》2024年第18期2721-2729,共9页Chongqing Medical Journal

基　　金：河北省自然科学基金项目(H2023209047/H2021209026);河北省人力资源和社会保障厅项目(C20210340);河北省重点研发计划项目(213777115D);河北省财政厅项目(冀财预复[2020]397号)。

摘　　要：目的探讨CXXC指蛋白4(CXXC4)对胰腺癌细胞PANC-1增殖和凋亡的影响。方法在线数据库分析胰腺癌组织中CXXC4的表达水平及其与患者预后和临床病理分期的关系。实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测人正常胰腺导管上皮细胞HPNE细胞及胰腺癌细胞PANC-1、AsPC-1和BxPC-3中CXXC4的mRNA表达。si-NC、si-CXXC4、pcDNA3.1和pcDNA3.1-CXXC4分别转染PANC-1细胞,Western bolt试验检测CXXC4敲降和过表达的有效性。CCK-8、集落形成、EdU和免疫荧光实验分析敲降或过表达CXXC4对PANC-1细胞增殖和凋亡的影响。生物信息学网站预测CXXC4的上游miRNA。Starbase数据库分析miR-450b-5p与CXXC4在胰腺癌组织中表达相关性。结果TCGA数据库结果显示,与癌旁胰腺组织相比,胰腺癌组织中CXXC4低表达(P<0.001),且与胰腺癌患者总体生存期和预后不良有关(P<0.05)。GEPIA数据库分析结果显示,与Ⅰ期胰腺癌相比,Ⅱ期胰腺癌中CXXC4表达降低(均P<0.05)。与HPNE细胞相比,在3种胰腺癌细胞中CXXC4表达降低(P<0.05)。与si-NC组相比,si-CXXC4组PANC-1细胞增殖和集落形成能力增强,增殖标志物Ki67、增殖细胞核抗原(PCNA)表达升高,凋亡标志物Bax、caspase-3及caspase-9表达降低;与pcDNA3.1组相比,pcDNA3.1-CXXC4组PANC-1细胞得到与上述相反的结果(均P<0.05)。生物信息学网站预测miR-450b-5p为CXXC4的上游miRNA,胰腺癌组织中CXXC4与miR-450b-5p的表达呈负相关(r=-0.227),miR-450b-5p在多种哺乳动物中物种保守较高。结论CXXC4抑制胰腺癌PANC-1细胞的增殖并促进其凋亡。Objective To investigate the effect of CXXC finger protein 4(CXXC4)on the proliferation and apoptosis of pancreatic cancer PANC-1 cells.Methods The expression level of CXXC4 in pancreatic cancer tissues and its relationship with prognosis and clinicopathological stage of the patients were analyzed in online databases.The qRT-PCR technique was used to detect the mRNA expression level of CXXC4 in human normal pancreatic ductal epithelial cell line(HPNE)and pancreatic cancer cell PANC-1,AsPC-1 and BxPC-3.si-NC,si-CXXC4,pcDNA3.1 and pCDNA3.1-CXXC4 were respectively transfected into PANC-1 cells.Western blot was conducted to detect the effectiveness of CXXC4 knockdown and overexpression.CCK-8,colony formation,EdU and immunofluorescence assays were conducted to analyze the effect of CXXC4 knockdown or overexpression on the proliferation and apoptosis of PANC-1 cells.The bioinformatic websites was used to predict the upstream microRNA(miRNA)of CXXC4.The Starbase database was adopted to analyze the correlation between miR-450b-5p and CXXC4 expression in pancreatic cancer tissues.Results The TCGA database results showed that the expression of CXXC4 in pancreatic cancer tissues was lowly expressed compared with in paracancerous pancreatic tissues(P<0.001),moreover which was associated with the overall survival and poor prognosis in the patients with pancreatic cancer(P<0.05).The GEPIA database analysis results showed that compared with stageⅠpancreatic cancer,the CXXC4 expression in stageⅡpancreatic cancer was decreased(P<0.05).Compared with HPNE cells,the CXXC4 expression in 3 kinds of pancreatic cancer cells was decreased(P<0.05).Compared with the si-NC group,the proliferation and colony formation ability of PANC-1 cells in the si-CXXC4 group were enhanced,the expressions of proliferation markers Ki67 and PCNA were increased,and the expressions of apoptosis markers Bax,caspase-3 and caspase-9 were decreased;compared with the pcDNA3.1 group,the PANC-1 cells in the pcDNA3.1-CXXC4 group obtained the opposite results(all

关 键 词：CXXC指蛋白4 胰腺癌 PANC-1细胞 增殖 凋亡 

分 类 号：R735.9[医药卫生—肿瘤]