敲低RBX1通过下调PHF5A抑制胰腺癌细胞增殖,迁移和侵袭并促进凋亡  

Knock-down RBX1 inhibits the proliferation,migration,and invasion of pancreatic cancer cells and promotes cell apoptosis by down-regulating PHF5A

作　　者：张菁 阎优优 徐栋超 张博 Zhang Jing;Yan Youyou;Xu Dongchao;Zhang Bo(Translational Medicine Research Center;Department of Gastroenterology,Affiliated Hangzhou First People’s Hospital,Zhejiang University School of Medicine)

机构地区：[1]浙江大学医学院附属杭州市第一人民医院转化医学研究中心,杭州310000 [2]浙江大学医学院附属杭州市第一人民医院消化内科,杭州310000

出　　处：《重庆医科大学学报》2023年第11期1314-1322,共9页Journal of Chongqing Medical University

基　　金：浙江省自然科学基金一般资助项目(编号:LY20H310005)。

摘　　要：目的:研究环盒蛋白1(ring-finger protein 1,RBX1)对胰腺癌(pancreatic adenocarcinoma,PAAD)细胞增殖、凋亡、迁移和侵袭的作用及其潜在的作用机制。方法:利用基因表达谱交互式分析网站(gene expression profile interactive analysis web⁃site,GEPIA),实时荧光定量聚合酶链反应(quantitative reverse transcription-polymerase chain reaction,qRT-PCR)、Western blot检测RBX1在PAAD组织、细胞中的表达和预后价值。使用细胞计数试剂盒-8(cell counting kit-8,CCK-8)、细胞克隆形成能力实验、脱氧核糖核苷酸末端转移酶介导的缺口末端标记法(terminal deoxynucleotidyl transferase dUTP nick end labeling,TUNEL)、细胞划痕实验、Transwell实验检测RBX1敲低对胰腺癌-1(pancreatic carcinoma-1,PANC-1)细胞增殖、凋亡、迁移和侵袭的影响。通过LinkedOmics网站研究PAAD中RBX1和PHD锌指结构域蛋白5A(plant homodomain finger-like domaincontaining protein 5A,PHF5A)的相关性。qRT-PCR和Western blot检测RBX1敲低后PHF5A的表达,进行功能拯救实验。结果:RBX1在PAAD组织和细胞系中高表达,与不良预后相关(P<0.05)。RBX1敲低抑制PANC-1细胞增殖、迁移和侵袭,促进细胞凋亡(P<0.05)。RBX1和PHF5A在PAAD中具有相关性(r=0.692),且RBX1正向调控PHF5A表达(P<0.05)。功能拯救实验表明PHF5A过表达部分逆转了RBX1敲低对PANC-1细胞增殖、迁移、侵袭和凋亡的作用(P<0.05)。结论:RBX1敲低通过下调PHF5A表达抑制PANC-1细胞增殖、迁移和侵袭,促进凋亡。Objective:To explore the effect of ring box protein-1(RBX1)on the proliferation,apoptosis,migration,and invasion of pancreatic adenocarcinoma(PAAD)cells and the potential mechanism of action.Methods:The gene expression profile interactive analysis website,quantitative real-time reverse-transcription polymerase chain reaction(qRT-PCR),and Western blot were used to measure the expression of RBX1 in PAAD tissues and cells and to determine its prognostic value.Cell Counting Kit-8,colony forma⁃tion assay,terminal deoxynucleotidyl transferase dUTP nick end labeling,wound healing assay,and transwell assay were applied to evaluate the effects of RBX1 knockdown on the proliferation,apoptosis,migration,and invasion of pancreatic carcinoma-1(PANC-1)cells.LinkedOmics website was used to investigate the correlation between RBX1 and plant homodomain finger-like domaincontaining protein 5A(PHF5A)in PAAD.Functional rescue experiments were performed by measuring the expression of PHF5A after RBX1 knockdown by qRT-PCR and Western blot.Results:RBX1 expression was up-regulated in PAAD tissues and cells,which was associated with a poor prognosis(P<0.05).RBX1 knockdown inhibited the proliferation,migration,and invasion of PANC-1 cells and promoted cell apoptosis(P<0.05).RBX1 was associated with PHF5A in PAAD(R=0.6918),and RBX1 positively regulated the expression of PHF5A(P<0.05).Functional rescue experiments showed that PHF5A overexpression partially reversed the effects of RBX1 knockdown on the proliferation,migration,invasion,and apoptosis of PANC-1 cells(P<0.05).Conclusion:RBX1 knockdown inhibits the proliferation,migration,and invasion of PANC-1 cells and promotes cell apoptosis by down-regulating PHF5A.

关 键 词：环盒蛋白1 PHD锌指结构域蛋白5A 胰腺癌 增殖 凋亡 侵袭迁移 

分 类 号：R730.23[医药卫生—肿瘤] R735.9[医药卫生—临床医学]