PTPN14通过调控Hippo/YAP信号通路抑制肾透明细胞癌细胞增殖和侵袭  

PTPN14 suppresses proliferation and invasion of clear cell renal carcinoma cells by regulating Hippo/YAP signaling pathway

作　　者：南宁[1] 刘莹[1] 强晓龙 马玉波 陈琦[1] NAN Ning;LIU Ying;QIANG Xiaolong;MA Yubo;CHEN Qi(Department of Urinary Surgery,Second Affiliated Hospital of Xi'an Jiaotong University,Xi’an 710004,China;Department of General Surgery, Zichang People's Hospital)

机构地区：[1]西安交通大学第二附属医院泌尿外科,西安710004 [2]子长市人民医院普外科

出　　处：《山西医科大学学报》2023年第7期910-916,共7页Journal of Shanxi Medical University

基　　金：陕西省自然科学基础研究计划项目(2023-JC-YB-649)。

摘　　要：目的探讨非受体酪氨酸蛋白磷酸酶14(non-receptor type protein tyrosine phosphatase 14,PTPN14)对肾透明细胞癌(clear cell renal cell carcinoma,ccRCC)细胞增殖和侵袭的调控作用及可能的作用机制。方法通过TCGA数据库分析PTPN14在ccRCC中的转录水平以及与生存率的关系。收集我院泌尿外科手术切除的ccRCC患者的肿瘤组织和癌旁正常组织,采用免疫组化检测PTPN14在ccRCC组织中的蛋白表达水平。采用实时定量PCR和Western blot检测PTPN14在正常肾小管上皮细胞和ccRCC细胞系中mRNA和蛋白表达水平。将Caki-1和786-O细胞分为对照组(转染空载体)和PTPN14组(转染PTPN14表达载体)。采用实时定量PCR检测PTPN14的mRNA表达水平,采用Western blot检测PTPN14、YAP、CTGF和CYR61的蛋白表达水平,运用EdU实验检测细胞增殖,运用Transwell实验检测细胞侵袭,运用荧光素酶报告基因实验检测YAP的转录活性。结果与正常组织比较,ccRCC组织中PTPN14转录水平显著降低(P<0.01)。高表达PTPN14的ccRCC患者总生存率显著高于低表达PTPN14的患者(P<0.01)。PTPN14在ccRCC组织中染色呈阴性。与正常肾小管上皮细胞比较,ccRCC细胞系中PTPN14的mRNA和蛋白表达水平显著减少(P<0.01)。与对照组比较,PTPN14组PTPN14的mRNA和蛋白表达水平显著升高,YAP、CTGF和CYR61的蛋白表达水平显著减少(均P<0.01)。与对照组比较,PTPN14组细胞增殖显著降低,细胞侵袭水平显著下降,YAP转录活性显著降低(均P<0.01)。结论PTPN14在ccRCC中表达水平降低,并通过调控Hippo/YAP信号通路抑制ccRCC细胞增殖和侵袭。Objective To explore the role of non-receptor type protein tyrosine phosphatase 14(PTPN14)in regulating the proliferation and invasion of clear cell renal carcinoma(ccRCC)cells and its potential molecular mechanism.Methods The transcript level of PTPN14 and its relationship with overall survival rate were evaluated based on the TCGA database.The protein expression of PTPN14 in tumor tissues and adjacent normal tissues from ccRCC patients undergoing surgery in Department of Urinary Surgery in our hospital was examined by the immumohistochemical staining method.The mRNA and protein expression levels of PTPN14 in normal renal tubular epithelial cells and ccRCC cell lines were detected by real-time quantitative PCR and Western blot.Caki-1 and 786-O cells were divided into control group and PTPN14 group,and transfected with empty vector and PTPN14 expression vector,respectively.The mRNA expression of PTPN14 was detected by real-time quantitative PCR.The protein expression of PTPN14,YAP,CTGF and CYR61 was detected by Western blot.Cell proliferation was assessed by EdU assay.Cell invasion was determined by Transwell assay.The transcriptional activity of YAP was measured by luciferase reporter assay.Results Compared with normal tissues,the transcript level of PTPN14 was significantly reduced in ccRCC tissues(P<0.01).The overall survival rate of ccRCC patients with high expression of PTPN14 was significantly higher than that of ccRCC patients with low expression of PTPN14(P<0.01).PTPN14 was negative in clinical tissues of ccRCC.Compared with normal renal tubular epithelial cells,PTPN14 mRNA and protein levels were significantly decreased in ccRCC cell lines(P<0.01).Compared with control group,the mRNA and protein levels of PTPN14 were significantly increased and the protein levels of YAP,CTGF,and CYR61 were significantly decreased in PTPN14 group(all P<0.01).Compared with control group,the cell proliferation was significantly reduced,the cell invasion was significantly decreased,and the transcriptional activity of YAP was

关 键 词：肾透明细胞癌 细胞增殖 细胞侵袭 PTPN14 YAP 

分 类 号：R737.11[医药卫生—肿瘤]