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

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 website，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 domain-containing 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。

Abstract:

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 formation 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 domain-containing 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.