胃癌患者外周血CD4+T细胞lncRNA心肌梗死转录本表达水平及其意义  

Expression and relevance of long non-coding RNA MIAT in CD4+T cells in peripheral blood of gastric cancer patients

作　　者：沙敏[1] 钱静宇 张传猛 程之逸[2] 朱雅斌[3] 金世光 Sha Min;Qian Jingyu;Zhang Chuanmeng;Cheng Zhiyi;Zhu Yabin;Jin Shiguang(Department of Clinical Laboratory,Taizhou People′s Hospital Affiliated of Nanjing Medical University,Taizhou 225300,China;Department of Gastrointestinal Surgery,Taizhou People′s Hospital Affiliated of Nanjing Medical University,Taizhou 225300,China;Department of Anesthesiology,Taizhou People′s Hospital Affiliated of Nanjing Medical University,Taizhou 225300,China)

机构地区：[1]泰州市人民医院中心实验室,泰州225300 [2]泰州市人民医院胃肠外科,泰州225300 [3]泰州市人民医院麻醉科,泰州225300

出　　处：《中华检验医学杂志》2022年第9期921-929,共9页Chinese Journal of Laboratory Medicine

基　　金：江苏省医学创新团队(CXTDA2017042);江苏省博士后资助项目(2019Z156);泰州市医学创学团队(CXTDB201904);泰州市人民医院院级课题(ZL201930)。

摘　　要：目的研究长链非编码RNA(lncRNA)心肌梗死转录本(MIAT)在胃癌患者外周血白细胞分化抗原(CD)4+T细胞中的表达变化及临床应用价值。方法收集江苏省泰州市人民医院2019年1月至2021年4月就诊的124例胃癌患者,90例胃良性疾病患者和80名健康对照者的外周血CD4+T细胞。分别应用实时荧光定量PCR和染色质免疫共沉淀联合定量聚合酶链反应检测MIAT表达量和RNA 6-甲基腺嘌呤(m6A)与MIAT基因启动子结合活性。用Spearman检验分析MIAT与临床病理特征、调节性T细胞(Treg)水平之间的相关性,用受试者工作特征(ROC)曲线评价CD4+T细胞中MIAT对胃癌的诊断效能。结果胃癌患者、胃良性疾病患者和健康对照者外周血CD4+T细胞中MIAT的相对表达量分别为2.849(2.131,4.062)、1.511(0.916,1.855)和0.963(0.729,1.432),3组间差异有统计学意义(H=158.25,P<0.001)。胃癌患者MIAT的表达水平显著高于胃良性疾病患者以及健康对照组,差异有统计学意义(Z=100.63,145.14,P均<0.001)。胃癌早期(Ⅰ~Ⅱ期)、晚期(Ⅲ~Ⅳ期)患者CD4+T细胞中m6A与MIAT基因启动子的相对结合活性分别为(8.590±1.483)和(4.274±0.425),差异有统计学意义(t=6.255,P=0.002);胃癌早、晚期患者m6A与MIAT基因的结合活性显著低于胃良性病变患者(17.267±3.106)和健康对照组(27.637±3.945)(t=-7.331、-12.832,P均<0.001)。胃癌患者MIAT相对表达量与年龄、性别、CEA水平和CA199水平无关(χ^(2)=0.000、0.000、0.648和1.554,P均>0.05),在不同TNM分期(χ^(2)=23.571,P=0.002)、不同肿瘤大小(χ^(2)=9.443,P<0.01)、有无淋巴结转移(χ^(2)=45.248,P<0.01)中差异有统计学意义。胃癌患者CD4+T细胞中MIAT相对表达量与Treg细胞比例之间呈正相关(r2=0.76,P<0.001)。用ROC曲线分析CD4+T细胞MIAT、血清CEA及CA199对胃癌患者的诊断效能,胃癌患者和胃良性疾病患者比较时,其曲线下面积分别为0.879、0.635和0.611;胃癌患者和健康者比较时,其曲线下面积Objective To explore the expression of long-chain noncoding RNA(lncRNA)and myocardial infarction-associated transcription(MIAT)in Leukocyte differentiation antigen(CD)4+T cells in peripheral blood of gastric cancer patients and its value of clinical application.Methods Peripheral blood CD4+T cells were collected from 124 patients with gastric cancer,90 benign gastric diseases patients and 80 healthy controls enrolled in Taizhou People′s Hospital from January 2019 to April 2021.The expression levels of MIAT and N6-methyladenosine(m6A)binding to MIAT promoter in CD4+T cells were detected by real-time fluorescent quantitative polymerase chain reaction(qPCR)and Chromatin immunoprecipitation(ChIP)-qPCR,respectively.Spearman test was used to analyze the correlation between MIAT and clinicopathological features,as well as between MIAT and regulatory T cell levels.The receivor operating characteristic curve(ROC)of the subjects was used to evaluate the MIAT expression level in the auxiliary diagnostic value of gastric cancer.Results The relative expression levels of MIAT in the gastric cancer patients,the benign gastric diseases patients,and the healthy controls were 2.849(2.131,4.062),1.511(0.916,1.855)and 0.963(0.729,1.432),respectively.The difference among the three groups was statistically significant(H=158.25,P<0.001).The relative expression level of MIAT in the gastric cancer patients was significantly higher than the levels in the benign gastric diseases patients and healthy controls.The difference was statistically significant(Z=100.63,145.14,P<0.001).The binding activity of m6A to MIAT promoter in patients with early stage(stageⅠandⅡ)and end stage(stageⅢandⅣ)gastric cancer was 8.590±1.483 and 4.274±0.425,respectively.The difference was statistically significant(t=6.255,P=0.002).Furthermore,the binding activity of m6A to MIAT promoter in the gastric cancer patients was significantly lower than that in patients with benign gastric diseases(17.267±3.106)and healthy controls(27.637±3.945)(t=-7.331,-12.8

关 键 词：胃肿瘤 CD4阳性T淋巴细胞 长链非编码RNA MIAT m6A 生物学标记 

分 类 号：R735.2[医药卫生—肿瘤]