转化生长因子β1对膀胱癌细胞增殖与迁移能力的影响及其机制  

EFFECT OF TRANSFORMING GROWTH FACTORβ1 ON PROLIFERATION AND MIGRATION ABILITY OF BLADDER CANCER CELLS AND ITS MECHANISM

作　　者：陈鑫磊 余永波 李鹏[1] 牛海涛[1] CHEN Xinlei;YU Yongbo;LI Peng;NIU Haitao(Department of Urology,The Affiliated Hospital of Qingdao University,Qingdao 266003,China)

机构地区：[1]青岛大学附属医院泌尿外科,山东青岛266003

出　　处：《精准医学杂志》2023年第2期111-115,共5页Journal of Precision Medicine

基　　金：国家自然科学基金面上项目(82071750);国家自然科学基金面上项目(81972378)。

摘　　要：目的探讨转化生长因子β1(TGF-β1)对膀胱癌T24细胞增殖、迁移能力的影响及其机制。方法以浓度为0、1、5、10、20、40μg/L的TGF-β1处理T24细胞24 h,采用MTT法检测细胞的增殖活性,采用细胞划痕方法检测其迁移能力,采用Western blot检测细胞中程序性死亡配体-1(PD-L1)蛋白的相对表达量;使用浓度为5μg/L的TGF-β1处理T24细胞0、0.5、1、2、6、24 h,采用Western blot实验检测细胞PD-L1蛋白及磷酸化丝苏氨酸蛋白激酶(p-AKT)和磷酸化细胞外调节激酶(p-ERK)蛋白的相对表达量;将T24细胞分为A~C组,A组使用正常培养基培养,B组培养基中加入终浓度为5μg/L的TGF-β1培养,C组培养基加入终浓度为5μg/L的TGF-β1及SB431542培养,Western blot方法检测A~C组细胞中PD-L1相对表达量;将T24细胞分为D~G组,D组使用正常培养基培养,E组培养基中加入终浓度为5μg/L的TGF-β1培养,F组培养基中加入终浓度为5μg/L的TGF-β1、AKT抑制剂MK2206培养,G组培养基加入终浓度5μg/L的TGF-β1、ERK抑制剂U0126处理T24细胞,均处理24 h,Western blot方法检测D~G组细胞中p-AKT、p-ERK和PD-L1相对表达量。结果浓度0、1、5、10、20、40μg/L的TGF-β1处理T24细胞24 h,其他浓度的TGF-β1处理T24细胞的增殖能力相较于0μg/L时均显著增强(t=3.59~12.18,P<0.05),各浓度之间T24细胞的迁移率没有显著差异(P>0.05),浓度为5μg/L时相较于其他浓度,PD-L1蛋白表达明显升高(t=3.22~5.64,P<0.05);浓度为5μg/L的TGF-β1处理T24细胞0、0.5、1、2、6、24 h后,T24细胞PD-L1蛋白表达随时间延长逐渐升高(F=199.20,P<0.05),并且处理时间为0.5 h相较于其他处理时间,p-AKT、p-ERK表达量显著升高(t=6.26~64.24,P<0.05);B组与A组比较,T24细胞PD-L1蛋白表达水平明显升高(t=31.24,P<0.05),C组与B组比较,PD-L1蛋白的表达水平降低(t=17.04,P<0.05);E组与D组比较,T24细胞的PD-L1、p-AKT、p-ERK蛋白水平明显升高(t=9.44~37.29,P<0.05),G组与E组比较,p-AKT、PD-L1�Objective To investigate the effect of transforming growth factorβ1(TGF-β1)on the proliferation and migration ability of bladder cancer T24 cells and its mechanism.Methods T24 cells were treated with TGF-β1 at concentrations of 0,1,5,10,20 and 40μg/L for 24 h.The proliferation activity of T24 cells was determined by methyl thiazolyl tetrazolium assay;their migration ability was detected by cell scratching method;the relative expression of programmed death ligand-1(PD-L1)protein in T24 cells was detected by Western blot.T24 cells were treated with TGF-β1 at a concentration of 5μg/L for 0,0.5,1,2,6 and 24 h.The relative expression of PD-L1 protein as well as phosphorylated serine-threonine protein kinase(p-AKT)and phosphorylated extracellular regulated kinase(p-ERK)proteins in T24 cells were detected by Western blot.T24 cells were divided into groups A to G.Group A was cultured with normal medium;group B was cultured with the medium containing TGF-β1 at a final concentration of 5μg/L;group C was cultured with the medium containing TGF-β1 and SB431542 at a final concentration of 5μg/L.The relative expression of PD-L1 in groups A to C was measured by Western blot.Group D was cultured with normal medium;group E was cultured with the medium containing TGF-β1 at a final concentration of 5μg/L;group F was cultured with the medium containing TGF-β1 and AKT inhibitor MK2206 at a final concentration of 5μg/L;group G was cultured with the medium containing TGF-β1 and ERK inhibitor U0126 at a final concentration of 5μg/L.All T24 cells were treated for 24 h.The relative expression of p-AKT,p-ERK,and PD-L1 proteins in groups D to G was detected by Western blot.Results After T24 cells were treated with TGF-β1 at concentrations of 0,1,5,10,20 and 40μg/L for 24 h,the proliferation ability of T24 cells was significantly enhanced compared with that of T24 cells treated with TGF-β1 at 0μg/L(t=3.59-12.18,P<0.05);the migration rate of T24 cells did not differ significantly between the concentrations(P>0.05);the PD-L

关 键 词：膀胱肿瘤 转化生长因子Β1 细胞增殖 细胞运动 B7-H1抗原 蛋白质丝氨酸苏氨酸激酶 丝裂原活化蛋白激酶3 

分 类 号：R737.14[医药卫生—肿瘤]