应激诱导磷酸化蛋白1调控GATA5表达促进结肠癌细胞增殖、侵袭和迁移过程中的作用及其机制  

Effects and mechanism of stress induced phosphoprotein 1 regulates GATA5 expression on proliferation,invasion and migration in colon cancer cells

作　　者：袁甲翔[1] 王佳辰[1] 王群[1] 沈新生[1] 王钊[1] 司亚卿[1] 翟二涛[2] Yuan Jiaxiang;Wang Jiachen;Wang Qun;Shen Xinsheng;Wang Zhao;Si Yaqing;Zhai Ertao(Department of Laparoscopic Surgery,the First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China;Department of Gastrointestinal Surgery,the First Affiliated Hospital of Sun Yat-Sen University,Guangzhou 510080,China)

机构地区：[1]郑州大学第一附属医院腹腔镜外科,450052 [2]中山大学附属第一医院胃肠外科中心,广州510080

出　　处：《中华实验外科杂志》2021年第11期2167-2170,共4页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金(82003112)。

摘　　要：目的检测应激诱导磷酸化蛋白1(STIP1)和GATA5在结肠癌细胞中的表达,探讨STIP1通过调节GATA5的表达在结肠癌细胞增殖、侵袭及迁移过程中的作用及机制。方法实时定量反转录聚合酶链反应(RT-qPCR)和蛋白质印迹法(Western blot)检测正常结肠(NCM460)和结肠癌(HCT116)2种细胞中STIP1、GATA5的mRNA及蛋白的表达。转染STIP1过表达载体和siRNA感染序列至HCT116细胞,实验分为3组,过表达转染组、阴性转染对照组、抑制组。Western blot检测STIP1、GATA5和β-连环蛋白(β-catenin)的表达。细胞计数试剂盒(CCK-8)、Transwell小室迁移实验和Matrigel侵袭实验检测细胞增殖、迁移及侵袭能力。采用t检验进行比较。结果 STIP1 mRNA和蛋白在HCT116的相对表达量分别为1.703±0.068、1.954±0.081,明显高于NCM460(1.000±0.000、1.000±0.000,t=17.78、20.236,P<0.01)。GATA5 mRNA和蛋白在HCT116的相对表达量分别为0.774±0.026、0.646±0.048,明显低于NCM460(1.000±0.000、1.000±0.000,t=14.76、12.673,P<0.01)。转染STIP1过表达载体后GATA5 mRNA和蛋白的相对表达量(0.534±0.065、0.742±0.058)明显低于对照组(1.000±0.000、1.000±0.000,t=12.27、7.621,P<0.01)。转染48 h和72 h后细胞增殖水平明显高于对照组(1.268±0.078比0.891±0.025,t=7.918,P<0.01;1.873±0.058比1.446±0.055,t=9.138,P<0.01);侵袭和迁移细胞数[(123.3±14.7)、(141.4±14.2)个]明显高于对照组[(94.9±14.2)、(100.8±12.1)个,t=4.369、6.852,P<0.01];β-catenin的表达(1.316±0.054)明显高于对照组(1.000±0.000,t=10.013,P<0.01]。转染STIP1 siRNA感染序列后GATA5 mRNA和蛋白的相对表达量(1.386±0.038、1.465±0.070)明显高于对照组(1.000±0.000、1.000±0.000,t=17.595、11.503,P<0.01)。转染72 h后细胞增殖水平(1.254±0.102)明显低于对照组(1.589±0.041,t=-5.243,P<0.01);侵袭和迁移细胞数[(60.8±8.7)、(77.0±7.9)个]明显低于对照组[(91.7±8.3)、(108.8±9.3)个,t=-8.088、-8.182,P<0.01];β-catenin的表达(0.741±0.046)�Objective To investigate the expression of stress induced phosphoprotein 1(STIP1)and GATA5,the effects and mechanism of STIP1 on proliferation,invasion and migration through regulating the expression of GATA5 in colon cancer cells.Methods The mRNA and protein expression levels of STIP1 and GATA5 were detected by real-time quantitative reverse transcriptase-polymerase chain reaction(RT-qPCR)and Western blotting in two cells(NCM460 and HCT116).The STIP1 over-expression vector and small interfering RNA(siRNA)were transfected into HCT116.The experiment was divided into three groups:overexpression transfection group,negative transfection control group and inhibition group.The expression of STIP1,GATA5 andβ-catenin were detected by Western blotting.Cell proliferation was detected by cell counting kit-8(CCK-8)assay,invasion and migration were detected by Transwell.The measurement data are expressed as mean±standard deviation and compared by t-test.Results The expression of STIP1 mRNA and protein in HCT116(1.703±0.068,1.954±0.081)were significantly higher than those in NCM460(1.000±0.000,1.000±0.000,t=17.78,20.236,P<0.01).The expression of GATA5 mRNA and protein in HCT116(0.774±0.026,0.646±0.048)were significantly lower than those in NCM460(1.000±0.000,1.000±0.000,t=14.76,12.673,P<0.01).The relative expression of GATA5 mRNA and protein were significantly lower(0.534±0.065 vs.1.000±0.000,t=12.27,P<0.01;0.742±0.058 vs.1.000±0.000,t=7.621,P<0.01),cell proliferation after transfected for 48 h and 72 h were significantly increased(1.268±0.078 vs.0.891±0.025,t=7.918,P<0.01;1.873±0.058 vs.1.446±0.055,t=9.138,P<0.01),the number of invasion and migration cells were significantly increased(123.3±14.7 vs.94.9±14.2,t=4.369,P<0.01;141.4±14.2 vs.100.8±12.1,t=6.852,P<0.01),β-catenin expression was higher(1.316±0.054 vs.1.000±0.000,t=10.013,P<0.01)in the STIP1 over-expression group.The relative expression of GATA5 mRNA and protein were significantly higher(1.386±0.038 vs.1.000±0.000,t=17.595,P<0.01;1.465±0.07

关 键 词：结肠癌 应激诱导磷酸化蛋白1 GATA5 Wnt-β-连环蛋白通路 

分 类 号：R735.35[医药卫生—肿瘤]