SRPK1促进肺腺癌进展的机制研究  

Mechanism of SRPK1 promoting lung adenocarcinoma progression

作　　者：焦敏 卢迪 刘泽毅[1] JIAO Min;LU Di;LIU Zeyi(Department of Pulmonary and Critical Care Medcine,First Affiliated Hospital of Soochow University,Suzhou 215006,China)

机构地区：[1]苏州大学附属第一医院呼吸与危重症医学科,苏州215006

出　　处：《山西医科大学学报》2024年第11期1381-1387,共7页Journal of Shanxi Medical University

基　　金：国家自然科学基金资助项目(82073213)。

摘　　要：目的探究丝氨酸-精氨酸蛋白激酶1(serine-arginine protein kinase 1,SRPK1)在肺腺癌中的表达及其促进肺腺癌恶性进展的机制。方法检索Human Protein Atlas数据库,比较SRPK1在正常肺组织和肺腺癌组织的表达水平;检索Kaplan Meier Plotter数据库,分析SRPK1的表达水平与肺腺癌患者生存预后的关系。采用慢病毒感染法在肺腺癌细胞A549、H1299中稳定过表达SRPK1,通过RT-qPCR和Western blot检测对照组和SRPK1过表达组细胞中SRPK1的表达水平。检索文献寻找SRPK1影响肿瘤进展的可能机制,通过Western blot检测对照组和SRPK1过表达组细胞中黏着斑激酶(focal adhesion kinase,FAK)及pFAK(Y397)的表达水平;检索癌症基因组图谱(The Cancer Genome Atlas,TCGA)数据库中肺腺癌队列的RNA-Seq数据,对SRPK1和丝氨酸蛋白酶抑制剂mRNA结合蛋白1(serpine 1 mRNA binding protein 1,SERBP1)的表达进行皮尔森相关性分析。通过细胞免疫荧光实验鉴定A549和H1299细胞中SRPK1和SERBP1的共定位情况;通过co-IP实验鉴定A549和H1299细胞中SRPK1和SERBP1的结合情况;采用慢病毒感染法在A549和H1299中稳定过表达SERBP1,通过RT-qPCR和Western blot检测对照组和SERBP1过表达组细胞中SERBP1的表达水平;通过RT-qPCR检测对照组和SERBP1过表达组细胞中SRPK1的表达水平;通过Western blot检测对照组和SERBP1过表达组细胞中SRPK1、FAK、pFAK(Y397)的表达水平。结果Human Protein Atlas数据库的数据显示,与正常肺组织相比,肺腺癌组织中SRPK1表达明显上调;Kaplan Meier Plotter数据库中数据显示,SRPK1表达水平越高,肺腺癌患者的预后越差(P=1.5×10^(-8))。RT-qPCR和Western blot结果显示,与对照组相比,SRPK1过表达组细胞SRPK1的表达水平上调(P<0.001);Western blot结果显示,与对照组相比,SRPK1过表达组细胞中FAK的表达基本不变,pFAK(Y397)的表达上调。皮尔森相关性分析显示,在TCGA数据库的肺腺癌队列数据中,SERBP1的基因表达与SRPK1的Objective To explore the expression of serine-arginine protein kinase 1(SRPK1)in lung adenocarcinoma and the mecha-nism of SRPK1 promoting lung adenocarcinoma progression.Methods The data from the Human Protein Atlas database were used to compare the expression of SRPK1 between normal lung tissues and lung adenocarcinoma tissues.The data from the Kaplan Meier Plot-ter database were used to analyze the relationship between SRPK1 expression and prognosis of lung adenocarcinoma patients.Lentivi-rus infection methods were applied to stably overexpress SRPK1 in lung adenocarcinoma cells A549 and H1299,and the expression of SRPK1 in control group and SRPK1 overexpression group was detected by RT-qPCR and Western blot.The possible mechanism of SRPK1 promoting tumor progression was firstly summarized from previous reports,and then the expressions of focal adhesion kinase(FAK)and pFAK(Y397)in control group and SRPK1 overexpression group were detected by Western blot.The RNA-Seq expression data of lung adenocarcinoma cohort in The Cancer Genome Atlas(TCGA)database were used to analyze the correlation between SRPK1 and serpine 1 mRNA binding protein 1(SERBP1)by Pearson correlation analysis.The colocalization of SRPK1 and SERBP1 in A549 and H1299 cells was identified by cell immunofluorescence assay.The binding of SRPK1 and SERBP1 in A549 and H1299 cells was verified by co-IP assay.Lentivirus infection methods were applied to stably overexpress SERBP1 in lung adenocarcinoma cells A549 and H1299,and the expression of SERBP1 in control group and SERBP1 overexpression group was detected by RT-qPCR and Western blot.The expression of SRPK1 in control group and SERBP1 overexpression group was detected by RT-qPCR.The expres-sion levels of SRPK1,FAK,and pFAK(Y397)in control group and SERBP1 overexpression group were detected by Western blot.Results The data from the Human Protein Atlas database showed that compared with normal lung tissue,the expression of SRPK1 in lung adenocarcinoma tissue was significantly upregulated.The data fr

关 键 词：肺腺癌 丝氨酸-精氨酸蛋白激酶1 黏着斑激酶 丝氨酸蛋白酶抑制剂mRNA结合蛋白1 恶性进展 预后 

分 类 号：R734.2[医药卫生—肿瘤]