SOX4通过靶向调节miR-17表达水平对结直肠癌细胞免疫逃逸及细胞迁移的影响  

Effect of SOX4 on immune escape and cell migration of colorectal cancer cells by targeting miR-17 expression level

作　　者：王国强 张纲 唐建坡 张玉国 杨永江[2] 王俊新 WANG Guoqiang;ZHANG Gang;TANG Jianpo;ZHANG Yuguo;YANG Yongjiang;WANG Junxin(Department of Gastrointestinal Surgery,the First Hospital of Zhangjiakou,Zhangjiakou,Hebei 075000,China;Department of Gastrointestinal tumor Surgery,the First Affiliated Hospital of Hebei North University,Zhangjiakou,Hebei 075000,China)

机构地区：[1]河北省张家口市第一医院胃肠外科,河北张家口075000 [2]河北北方学院附属第一医院胃肠肿瘤外科,河北张家口075000

出　　处：《检验医学与临床》2024年第13期1825-1830,1835,共7页Laboratory Medicine and Clinic

基　　金：国家自然科学基金资助项目(19NSP045);河北省医学科学研究课题计划(20211329)。

摘　　要：目的探究性别决定区Y框蛋白4(SOX4)通过靶向调节微小RNA(miR)-17表达水平对结直肠癌细胞免疫逃逸及细胞迁移的影响。方法采用实时荧光定量聚合酶链反应检测正常结直肠黏膜上皮细胞及4种结直肠癌细胞中SOX4、miR-17表达水平。取结直肠癌细胞将其分为结直肠癌细胞(CC)组、结直肠癌细胞+SOX4-NC(SN)组、结直肠癌细胞+SOX4激动剂(SM)组、结直肠癌细胞+SOX4抑制剂(SI)组、结直肠癌细胞+miR-17-NC(MN)组、结直肠癌细胞+miR-17激动剂(MM)组、结直肠癌细胞+miR-17抑制剂(MI)组、结直肠癌细胞+SOX4抑制剂+miR-17抑制剂(II)组。采用免疫印迹法检测程序性死亡受体-1(PD-1)及PD-1配体(PD-L1)表达水平,小室法检测细胞迁移数量,双荧光素酶报告基因检测各组细胞荧光素酶活性。结果4种结直肠癌细胞中SOX4、miR-17 mRNA表达水平明显高于FHC正常结直肠黏膜上皮细胞,差异均有统计学意义(P<0.05)。CC组与SN组PD-1、PD-L1蛋白表达水平及细胞迁移数量比较,差异均无统计学意义(P>0.05)。SM组PD-1、PD-L1蛋白表达水平及细胞迁移数量均明显高于SN组,SI组PD-1、PD-L1蛋白表达水平及细胞迁移数量均明显低于SM组,差异均有统计学意义(P<0.05)。CC组与MN组PD-1、PD-L1蛋白表达水平及细胞迁移数量比较,差异均无统计学意义(P>0.05)。MM组PD-1、PD-L1蛋白表达水平及细胞迁移数量均明显高于MN组,MI组PD-1、PD-L1蛋白表达水平及细胞迁移数量均明显低于MM组,差异均有统计学意义(P<0.05)。SI组与MI组PD-1、PD-L1蛋白表达水平及细胞迁移数量比较,差异均无统计学意义(P>0.05)。II组PD-1、PD-L1蛋白表达水平及细胞迁移数量均明显低于MI组,差异均有统计学意义(P<0.05)。双荧光素酶报告基因检测结果显示,转染SOX4后miR-17-3′-UTR-WT的荧光素酶活性明显升高,差异有统计学意义(P<0.05),miR-17-3′-UTR-MUT的荧光酶活性无明显变化,差异无统计学意义(P>0.Objective To investigate the effect of sex determining region Y-box 4(SOX4)on immune escape and cell migration of colorectal cancer cells by targeting microrna(miR)-17 expression.Methods The expression levels of SOX4 and miR-17 in normal colorectal mucosal epithelial cells and 4 types of colorectal cancer cells were detected by real-time fluorescence quantitative polymerase chain reaction.Take the colorectal cancer cell was divided into colorectal cancer cell(CC)+SOX4-NC group,the colorectal cancer cells(SN),colorectal cancer cells+SOX4 agonist(SM),colorectal cancer cells+SOX4 inhibitors(SI),colorectal cancer cell+miR-17-NC(MN)group,colorectal cancer cells+miR-17 agonist(MM)group,colorectal cancer cells+miR-17 inhibitor(MI)group,colorectal cancer cells+SOX4 inhibitor+miR-17 inhibitor(II)group.The expression levels of programmed death-1(PD-1)and PD-1 ligand 1(PD-L1)were detected by Western blot.The number of cell migration was detected by chamber method.The luciferase activity of cells in each group was detected by dual luciferase reporter gene.Results The expression levels of SOX4 and miR-17 mRNA in 4 kinds of colorectal cancer cells were significantly higher than those in FHC normal colorectal mucosal epithelial cells,and the differences were statistically significant(P<0.05).There was no significant difference in the expression level of PD-1 and PD-L1 protein and the number of cell migration between CC group and SN group(P>0.05).The expression levels of PD-1 and PD-L1 protein and the number of cell migration in SM group were significantly higher than those in SN group,and the expression levels of PD-1 and PD-L1 protein and the number of cell migration in SI group were significantly lower than those in SM group(P<0.05).There was no significant difference in the expression level of PD-1 and PD-L1 protein and the number of cell migration between CC group and MN group(P>0.05).The expression levels of PD-1 and PD-L1 protein and the number of cell migration in MM group were significantly higher than those in MN group

关 键 词：性别决定区Y框蛋白4 微小RNA-17 结直肠癌 免疫逃逸 细胞迁移 

分 类 号：R735.35[医药卫生—肿瘤] R446.1[医药卫生—临床医学]