质子泵抑制剂通过下调细胞周期相关基因诱导胃癌细胞周期阻滞并促进化疗增敏  

Proton Pump Inhibitors Induce Cell Cycle Arrest and Enhance Chemosensitivity of Gastric Cancer Cells by Inhibiting Cell Cycle-related Genes

作　　者：苏萌 张斌 陈敏[2] 王雷[1] 徐桂芳[1] 吕瑛[1] 邹晓平[1] SU Meng;ZHANG Bin;CHEN Min;WANG Lei;XU Guifang;L Ying;ZOU Xiaoping(Department of Gastroenterology,Nanjing Drum Tower Hospital,Clinical College of Nanjing Medical University,Nanjing,210008;Department of Gastroenterology,Nanjing Drum Tower Hospital,the Affiliated Hospital of Nanjing University Medical School,Nanjing)

机构地区：[1]南京医科大学鼓楼临床医学院,210008 [2]南京大学医学院附属鼓楼医院消化内科

出　　处：《胃肠病学》2021年第9期519-525,共7页Chinese Journal of Gastroenterology

基　　金：南京市医学科技发展项目杰出青年基金项目(JQX15001)。

摘　　要：背景:研究发现质子泵抑制剂(PPI)可增强胃癌细胞的化疗敏感性,抑制胃癌细胞增殖和侵袭。目的:探讨PPI是否系通过影响细胞周期相关基因表达对胃癌细胞发挥化疗增敏作用。方法:以不同浓度泮托拉唑处理人胃癌细胞株AGS和HGC27,CCK-8实验检测细胞活力,转录组测序结合KEGG富集分析明确PPI对胃癌细胞周期的影响,流式细胞分析、real-time PCR和蛋白质印迹法验证细胞周期及其相关基因表达变化。利用生物信息学网站分析主要细胞周期相关差异表达基因在胃癌组织中的表达及其与患者预后的关系。CCK-8实验检测PPI联合顺铂对转染FOXM1过表达质粒或空载质粒的胃癌细胞的增殖抑制效果。结果:PPI可有效抑制胃癌细胞体外增殖。转录组测序和KEGG富集分析显示经PPI处理的胃癌细胞G2/M期相关基因FOXM1、PLK1、AURKB等表达下调,并出现G2/M期阻滞,上述发现经流式细胞分析以及real-time PCR和蛋白质印迹法证实。生物信息学分析显示上述G2/M期相关基因在胃癌组织中的表达显著上调,并与预后不良相关。PPI联合顺铂对胃癌细胞的增殖抑制作用显著强于顺铂单药处理,但可被过表达FOXM1部分逆转。结论:PPI可通过下调细胞周期相关基因表达诱导G2/M期阻滞,从而增强胃癌细胞的化疗敏感性。Background:Several studies have shown that proton pump inhibitors(PPIs)can enhance the sensitivity of gastric cancer(GC)cells to chemotherapy and inhibit tumor proliferation and invasion.Aims:To investigate whether PPI could enhance chemosensitivity by inhibition of cell cycle-related genes in GC cells.Methods:Two human GC cell lines,AGS and HGC27 were treated with pantoprazole in different concentrations,and the cell viability was detected by CCK-8 assay.Transcriptome sequencing combined with KEGG enrichment analysis were used to determine the effect of PPI on cell cycle of GC cells,and the changes of cell cycle and its related genes were validated by flow cytometry,real-time PCR and Western blotting,respectively.Bioinformatics websites were employed to analyze the major differentially expressed cell cycle-related genes in GCs and their relationship with patients’prognosis.After transfection with FOXM1 plasmid or control plasmid,the inhibitory effect of PPI combined with cisplatin on GC cells was determined by CCK-8 assay.Results:PPI inhibited the proliferation of GC cells effectively in vitro.Transcriptome sequencing showed that the expression levels of G2/M phase-related genes,including FOXM1,PLK1,and AURKB were down-regulated in PPI-treated GC cells,and G2/M arrest was suggested by KEGG enrichment analysis.All these changes were proved by flow cytometry,real-time PCR and Western blotting.Bioinformatics analysis revealed that FOXM1,PLK1,and AURKB genes were highly expressed in GCs and correlated with a poor prognosis.The inhibitory effect of PPI combined with cisplatin on GC cells was superior to that of cisplatin alone,but could be partially reversed by overexpression of FOXM1.Conclusions:PPI treatment can induce G2/M arrest in GC cells by inhibiting cell cycle-related genes,and subsequently enhance the sensitivity of GC cells to chemotherapy.

关 键 词：胃肿瘤 质子泵抑制剂 细胞周期 G2/M期阻滞 化疗敏感性 

分 类 号：R735.2[医药卫生—肿瘤]