摘要
目的:探究催乳素受体(prolactin receptor,PRLR)在胰腺导管腺癌(pancreatic ductal adenocarinoma,PDAC)发生和发展过程中表达水平的变化,及其表达水平改变对PDAC细胞对耐药和凋亡的影响和可能的作用机制。方法:利用基因综合表达(Gene Expression Omnibus,GEO)数据库分析PRLR在PDAC及其癌旁组织中的表达水平及在肿瘤发生和发展过程中表达量的变化。癌症基因组图谱(e Cancer Genome Atlas,TCGA)分析PRLR表达水平与患者预后的相关性。采用实时荧光定量PCR、蛋白质印迹法和免疫组织化学法检测PRLR mRNA和蛋白在PDAC细胞和组织内的表达水平。同时,利用基因集富集分析(Gene Set Enrichment analysis,GSEA)等生物信息学手段推断PRLR在PDAC中可能涉及的信号通路。利用shRNA技术沉默PANC-1和SW-1990细胞中PRLR的表达水平后,分别采用CCK-8和FCM法检测下调PRLR表达对PDAC细胞对吉西他滨(gemcitabine)耐药和凋亡的影响,以及对耐药相关因子转酮醇酶(transketolase,TKT)和葡萄糖-6-磷酸脱氢酶(glucose-6-phosphatedehydrogenase,G6PD)表达的影响。结果:GEO数据分析结果表明,PRLR在PDAC组织中表达量下降。TCGA分析结果表明,PRLR低表达与患者的不良生存相关。GSEA分析结果表明,PRLR的表达量与PDAC细胞中Wnt通路、转化生长因子β(transforminggrowthfactor-β,TGF-β)通路和Janus激酶(janus kinase,JAK)-信号转导及转录激活因子(signaltransducerandactivator oftranscription,STAT)通路等耐药通路相关。基因共表达分析数据分析也表明,PRLR表达量的降低与肿瘤耐药基因谷胱甘肽-S-转移酶P1(glutathione-S-transferase P1,GSPT1)表达量升高相关,同时与抑制肿瘤耐药的黏附G蛋白偶联受体V1(adhesion G-protein coupled receptor V1,GPR98)表达量降低相关。敲低PANC-1和SW-1990细胞中PRLR的表达水平后,能够提高肿瘤细胞对于吉西他滨的耐药性并降低细胞的凋亡率(P均<0.01),并能够提高耐药相关因子TKT和G6PD的表达水平(P均<0.01)。结论:PRLR在PDAC中低表达,沉默PDAC细胞中PRLR的表达水平可进一步提高PDAC细胞的耐药性并降低细胞的凋亡能力,提示其可能是一个潜在的肿瘤治疗靶点。
Objective:To investigate the changes of prolactin receptor(PRLR)expression level during development of pancreatic ductal adenocarcinoma(PDAC),and its potential mechanism of drug resistance and apoptosis of PDAC cells.Methods:The gene expression of PRLR was analyzed in PDAC and normal pancreatic tissue during tumorigenesis and development with Gene Expression Omnibus(GEO)database.The expression and prognosis of PRLR was analyzed by The Cancer Genome Atlas(TCGA)database.The expression level of PRLR mRNA and protein level in PDAC and normal pancreatic tissue,the quantitative real time PCR,western blotting and immunohistochemistry were used.Meanwhile,the potential pathway was discovered with Gene Set Enrichment analysis(GSEA)and other bioinformatics method.The drug resistance of gemcitabine and apoptosis to PDAC cells were detected by CCK-8 and FCM respectively with the PANC-1 and SW-1990 cells line knockdown PRLR expression with short hairpin RNA(shRNA).Lastly,the expression relationship between PRLR and transketolase(TKT)and glucose-6-phosphatedehydrogenase(G6PD)was investigated in PDAC cells.Results:The result of GEO and TCGA data demonstrated that PRLR expression declined in PDAC tissue and correlated with patient poor prognosis respectively.In GESA analysis,the PRLR expression was discovered that related to Wnt,transforming growth factor-β(TGF-β),janus kinase(JAK)-signal transducer and activator of transcription(STAT)pathway which related to PDAC drug resistance.The gene co-expression analysis show that PRLR expression positively related to glutathione-S-transferase P1(GSPT1)which increased PDAC cells drug resistance and negatively related to adhesion G-protein coupled receptor V1(GPR98)which decreased PDAC cells drug resistance.In PANC-1 and SW-1990 cells,knockdown PRLR expression increased drug resistance to gemcitabine and decreased the apoptosis rate,and increased TKT and G6PD expression levels(all P<0.01).Conclusion:Depletion PRLR,which low express in PDAC tissue,will increased PDAC cell drug resistance and decreased PDAC cell apoptosis,which suggested that it may be a potential tumor therapeutic target.
作者
张一帆
黄佩琦
孙悦
潘泓
盖严支
聂惠贞
ZHANG Yifan;HUANG Peiqi;SUN Yue;PAN Hong;GAI Yanzhi;NIE Huizhen(State Key Laboratory of Oncogenes and Related Genes,Ren Ji Hospital,Shanghai Jiao Tong University of Medicine,Shanghai Jiao Tong University,Shanghai 200240,China)
出处
《肿瘤》
CAS
CSCD
北大核心
2021年第6期383-393,共11页
Tumor
基金
上海市卫健委资助课题(201740105)
关键词
胰腺导管腺癌
催乳素受体
耐药性
吉西他滨
Ductal adenocarcinoma of the pancreas
Prolactin receptor
Drug resistance
Gemcitabine
作者简介
Correspondence to:NIE Huizhen(聂惠贞),E-mail:hznie@shsci.org
