摘要
目的探讨外泌体miR-218-5p在结直肠癌(CRC)患者血清中的表达水平及其与患者临床病理特征的相关性,评价其在CRC中的诊断效能。方法选择同济大学附属同济医院2016年10月至2018年10月确诊的结直肠癌病例组78例,术前采血并保存血清;选择同期健康体检者40例作为对照组。采用ExoQuick试剂盒提取血清外泌体,应用透射电镜、NTA、蛋白印迹对外泌体进行形态学和分子表型鉴定;利用miRNeasy试剂盒提取血清外泌体中总RNA,实时荧光定量PCR法(RT-qPCR)检测各组血清外泌体miR-218-5p的表达水平。以miR-218-5p相对表达水平的中位数为截断值,将CRC患者分为高表达与低表达组,用χ^(2)检验判断其与临床病理特征的关系;受试者工作特征(ROC)曲线判断其在结直肠癌中的诊断效能。结果试剂盒法成功提取到血清中的外泌体。结直肠癌组血清外泌体miR-218-5p的表达水平明显低于对照组[0.566(0.364,0.850)比1.054(0.781,1.709),P<0.001];其低表达程度与肿瘤大小、TNM分期、肿瘤淋巴结转移和浸润深度显著相关(P均<0.05);ROC分析表明血清外泌体miR-218-5p诊断结直肠癌的曲线下面积(AUC)为0.827(95%CI 0.754~0.900),明显优于常规的肿瘤标志物癌胚抗原CEA(AUC=0.718,95%CI 0.626~0.811)和糖类抗原CA199(AUC=0.661,95%CI 0.564~0.758)。结论结直肠癌患者血清外泌体中miR-218-5p的表达下调与多个不良临床病理学因素相关,具有成为结直肠癌诊断生物标志物的潜力。
Objective To explore the expression level of exosomal miR-218-5p in the serum of patients with colorectal cancer(CRC)and its correlation with the clinical pathological characteristics,and evaluate its diagnostic efficacy in CRC.Methods A group of 78 patients with colorectal cancer diagnosed in Tongji Hospital affiliated to Tongji University from October 2016 to October 2018 were selected.Blood was collected before operation and serum was preserved.Forty cases of healthy people in the same period were selected as the control group.ExoQuick kit was used to extract serum exosomes.Transmission electron microscope,NTA and western blot were used to identify the morphology and molecular phenotype of exosomes.MiRNeasy kit was used to extract total RNA in serum exosomes.Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expression level of serum exosomal miR-218-5p in each group.CRC patients were divided into high expression and low expression groups using the median relative expression level of miR-218-5p as the cut off value,and the four-square chi-square test(χ^(2) test)was used to judge the relationship between miR-218-5p and clinicopathological features.The receiver operating characteristic(ROC)curve was used to evaluate the diagnostic efficacy of miR-218-5p in colorectal cancer.Results The exosomes in serum were successfully extracted by kit method.The expression level of serum exosomal miR-218-5p in patients with colorectal cancer was significantly lower than that in normal healthy people[0.566(0.364,0.850)vs 1.054(0.781,1.709),P<0.001].The low expression level was significantly better then the correlated with tumor size,TNM stage,lymph node metastasis and depth of invasion(all P<0.05).Receiver operating characteristic(ROC)analysis showed that the area under the curve(AUC)of serum exosomal miR-218-5p for the diagnosis of CRC was 0.827(95%CI 0.754-0.900),which was significantly better than the conventional tumor marker carcinoembryonic antigen CEA(AUC=0.718,95%CI 0.626-0.811)and carbohydrate anti
作者
常文婧
商安全
杨滇宇
田佳乐
孙祖俊
李冬
权文强
Chang Wenjing;Shang Anquan;Yang Dianyu;Tian Jiale;Sun Zujun;Li Dong;Quan Wenqiang(Department of Laboratory Medicine,Tongji Hospital,University of Tongji,Shanghai 200065,China)
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2021年第8期709-714,共6页
Chinese Journal of Laboratory Medicine
基金
国家自然科学基金(81902984,81873975)
上海市医疗卫生优秀学科带头人培养计划(2018BR31)
上海市科委医学引导项目(19411964800)
中国博士后科学基金(2020M681399)
同济大学附属同济医院优秀后备人才计划(HBRC2005)。
