乳腺癌中Notch信号通路与YAP1/TAZ的交互作用研究  

Correlation between Notch signaling pathway and YAP1/TAZ in breast cancer

作　　者：赵琳[1] 张玉姣[1] 雷福茜 罗安琪 景鑫[1] 王璐 雷建军[4] 邵珊[1] ZHAO Lin;ZHANG Yujiao;LEI Fuxi;LUO Anqi;JING Xin;WANG Lu;LEI Jianjun;SHAO Shan(Department of Oncology,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061;Department of Nephrology,The Second Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710004;Department of Nuclear Medicine,Cancer Hospital of Chinese Academy of Medical Sciences,Beijing 100021;Department of Hepatobiliary Surgery,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an 710061,China)

机构地区：[1]西安交通大学第一附属医院肿瘤内科,陕西西安710061 [2]西安交通大学第二附属医院肾脏内科,陕西西安710004 [3]中国医学科学院肿瘤医院核医学科,北京100021 [4]西安交通大学第一附属医院肝胆外科,陕西西安710061

出　　处：《西安交通大学学报（医学版）》2020年第6期888-895,共8页Journal of Xi’an Jiaotong University（Medical Sciences）

基　　金：国家自然科学基金资助项目(No.81502066,No.81702908);陕西省自然科学基础研究计划一般项目(2019JM-115,2018JQ8030);西安交通大学基础科研业务费资助(xzy012019090)。

摘　　要：目的探讨Notch1与YAP1/TAZ在乳腺癌中的表达关系及其发病机制。方法选取MDA-MB-231和MCF-7乳腺癌细胞,利用慢病毒分别构建不同Notch1、YAP1表达水平的稳定感染细胞株,应用Western blotting、RT-qPCR、免疫共沉淀技术研究Notch1与YAP1/TAZ之间的关系。结果通过基因表达谱交互分析(gene expression profiling interactive analysis,GEPIA),对肿瘤基因图谱数据库(the Cancer Genome Atlas,TCGA)中的乳腺癌数据分析发现,Notch1与YAP1/TAZ之间表达呈正相关;与对照组相比,shNotch1组YAP1/TAZ的蛋白水平与mRNA水平均降低,差异具有统计学意义(P<0.05),N1ICD组YAP1/TAZ的蛋白表达水平升高,mRNA水平差异无统计学意义(P>0.05),在Notch1过表达的MDA-MB-231细胞中,TAZ降解被抑制;免疫共沉淀实验结果显示,TAZ蛋白与Notch1、β-TrCP蛋白存在相互作用;与对照组相比,shYAP1组Notch1、JAG1的mRNA水平及蛋白水平均降低(P<0.05),YAP1组Notch1、JAG1的mRNA水平及蛋白水平均升高,差异具有统计学意义(P<0.05);JASPAR 2018网站预测TEAD家族为JAG1的转录因子;当siTEAD1/3/4#1、#2有效抑制TEAD1/3/4mRNA的转录与蛋白表达时,JAG1的mRNA和蛋白水平均降低(P<0.05)。结论在乳腺癌中,Notch1与YAP1/TAZ存在正反馈环路。YAP1/TAZ可能通过与TEAD家族结合调节JAG1的表达,活化Notch1信号通路。Notch1蛋白影响YAP1/TAZ蛋白的降解。Objective To investigate the relationship of Notch1 expression with YAP1/TAZ expression in breast cancer and its possible mechanism.Methods MDA-MB-231 and MCF-7 breast cancer cells were selected,and lentiviruses were used to construct stable infected cell lines with different expression levels of Notch1 and YAP1.Western blotting,RT-qPCR and co-immunoprecipitation were used to study the relationship between Notch1 and YAP1/TAZ expressions.Results On the gene expression profiling interactive analysis(GEPIA)website,we analyzed the breast cancer data in the Cancer Gene Atlas Database(TCGA)and found that the expressions of Notch1 and YAP1/TAZ were positively correlated.Compared with those in the control group,the protein and mRNA levels of YAP1/TAZ in the shNotch1 group were reduced(P<0.05).In N1ICD group,YAP1/TAZ protein expression level was increased,but there was no significant difference in mRNA level(P>0.05).Co-immunoprecipitation showed that TAZ protein interacted with Notch1 andβ-TrCP protein.Compared with those in the control group,the mRNA and protein levels of Notch1 and JAG1 in shYAP1 group were reduced(P<0.05),while those in YAP1 group were increased(P<0.05).TEAD family was predicted to be a JAG1 transcription factor on the JASPAR 2018 website.TEAD1/3/4 siRNA could effectively inhibit TEAD1/3/4 expression,and JAG1 expression decreased too(P<0.05).Conclusion There is a positive feedback loop between Notch1 and YAP1/TAZ in breast cancer.YAP1/TAZ-TEAD can activate the Notch1 signaling pathway by regulating JAG1 expression.Notch1 protein can affect the degradation of YAP1/TAZ protein.

关 键 词：乳腺癌 NOTCH1 YAP1/TAZ TEAD 

分 类 号：R737.9[医药卫生—肿瘤]