miR-4484调控整合素α 6在胃癌组织中的表达及其意义  

MiR-4484 regulates the expression of integrinα6 in gastric cancer tissues and its significance

作　　者：钟慧钰 钟源 温阳 陶昕彤 宋兴勃[1] 陆小军[1] Zhong Huiyu;Zhong Yuan;Wen Yang;Tao Xintong;Song Xingbo;Lu Xiaojun(Department of Experimental Medicine,West China Hospital,Sichuan University,Chengdu 610041,China)

机构地区：[1]四川大学华西医院实验医学科,成都610041

出　　处：《中华肿瘤杂志》2022年第3期246-251,共6页Chinese Journal of Oncology

基　　金：国家自然科学基金面上项目(81672096);四川省科技厅重点项目(2017FZ0065)。

摘　　要：目的探讨整合素α6(ITGA6)蛋白、基因和miR-4484与胃癌病理分期的关系。方法收集2017年6—9月于四川大学华西医院行手术治疗(未经辅助治疗)的30例原发性胃癌患者的胃癌及癌旁(距离瘤体>5 cm)正常胃黏膜组织。采用实时荧光定量PCR检测miR-4484和ITGΑ6的表达水平,采用Western blot法检测ITGA6蛋白的表达水平,采用双荧光素酶报告基因验证ITGΑ6与miR-4484的关系,采用Spearman相关分析明确miR-4484与ITGA6在胃癌组织中表达水平的相关性。结果胃癌组织中ITGA6的表达水平(32.30±13.47)高于正常癌旁组织(24.55±10.25,P=0.015),受试者工作特征(ROC)曲线下面积为0.660,其诊断敏感度为43.3%,特异度为96.7%。胃癌组织中miR-4484的表达水平(4.11±2.87)低于正常癌旁组织(5.75±2.80,P=0.029),ROC曲线下面积为0.690,其敏感度为30.0%,特异度为86.7%。胃癌组织中miR-4484表达水平与ITGA6呈负相关(r=-0.621,P<0.001)。ITGA6在胃癌组织中的表达水平(0.65±0.19)高于正常癌旁组织(0.26±0.12,P<0.001)。与ITGΑ63′UTR野生型+miR-NC组比较,ITGΑ63′UTR野生型+miRNA mimics组细胞荧光素酶活性降低(分别为50.69±5.10和34.00±1.19,P<0.001),ITGΑ63′UTR野生型+ASO miR-4484组细胞荧光素酶活性高于ITGΑ63′UTR野生型+miR-NC组(分别为82.44±6.37和50.69±5.10,P<0.001),ITGA6为miR-4484的直接靶基因。T1、T2、T3、T4(4a和4b)期胃癌组织中miR-4484的表达水平分别为9.98±2.24、5.28±2.03、2.92±2.04和4.11±2.87,差异有统计学意义(P<0.001)。N0、N1、N2、N3期胃癌组织中ITGA6的表达水平分别为29.55±8.32、21.71±3.75、24.60±8.79和40.69±15.83,差异有统计学意义(P=0.022)。N0、N1、N2、N3期胃癌组织中miR-4484的表达水平分别为5.01±3.52、5.48±2.76、5.88±1.83和2.30±1.56,差异有统计学意义(P=0.032)。M0、M1期胃癌组织中ITGA6的表达水平分别为26.28±7.66和52.08±8.12,差异有统计学意义(P<0.001)。M0、M1期胃癌组织中miR-4484的表达Objective To investigate the relationship between the expression of integrinα6(ITGA6),miR-4484 and the pathologic stage of gastric cancer.Methods Gastric cancer tissues and normal gastric mucosa tissues adjacent to cancer(>5 cm from tumor margin)of 30 patients with primary gastric cancer who underwent direct surgical resection without adjuvant therapy from June to September 2017 in West China Hospital of Sichuan University were selected.Real-time quantitative polymerase chain reaction(PCR)was used to detect the expression levels of miR-4484 and ITGA6,western blot was used to detect the expression level of ITGA6 protein,dual luciferase reporter gene was used to verify the relationship between ITGA6 and miR-4484.Spearman′s correlation analysis was used to determine the relationship between miR-4484 and ITGA6 expression levels in gastric cancer tissues.Results The expression level of ITGΑ6 in gastric cancer(32.30±13.47)was higher than that in matched normal gastric tissues(24.55±10.25,P=0.015),the area under the receiver operating characteristic(ROC)curve was 0.660 and the diagnostic sensitivity and specificity were 43.3%and 96.7%,respectively.The expression level of miR-4484 in gastric cancer(4.11±2.87)was lower than that of matched normal gastric tissues(5.75±2.80,P=0.029),the area under the ROC curve was 0.690 and the diagnostic sensitivity and specificity were 30.0%and 86.7%,respectively.The expression level of miR-4484 was negatively correlated with ITGA6 in gastric cancer tissues(r=-0.621,P<0.001).The expression level of ITGA6 protein in gastric cancer tissues(0.65±0.19)was higher than that in normal adjacent tissues(0.26±0.12,P<0.001).Compared with ITGA63′UTR wild-type+miR-NC group,ITGA63′UTR wild-type+miRNA mimics group had lower luciferase activity(50.69±5.10,34.00±1.19,P<0.001),while the luciferase activity of ITGA63′UTR wild-type+ASO miR-4484 group was higher than that of ITGA63′UTR wild-type+miR-NC group(82.44±6.37,50.69±5.10,P<0.001),indicated that ITGA6 was the direct target gene

关 键 词：胃肿瘤 整合素 MICRORNA 侵袭与转移 

分 类 号：R735.2[医药卫生—肿瘤]