摘要
目的探讨胰腺癌(PC)血清外泌体人膜联蛋白A11(ANXA11)的检测方法,初步评估ANXA11在PC中的临床应用价值。方法选择2016年12月至2019年7月南通大学附属医院确诊为PC、胰腺良性占位、胰腺炎患者的血清标本分别70例、15例、70例,选取同期健康体检者70名作为对照组。利用基于高分辨率、高精度质谱仪的平行反应监测(PRM)方法检测血清外泌体及无外泌体血清中ANXA11蛋白丰度。自建检测人血清外泌体ANXA11的点免疫印迹法(dot immunoblotting)并进行方法学评价;分析所有受试者血清外泌体ANXA11的水平;采用受试者工作特征(ROC)曲线的曲线下面积(AUC)评价ANXA11、糖类抗原19-9(CA19-9)、癌胚抗原(CEA)对PC的诊断效能;分析ANXA11与PC患者临床病理参数、疗效及预后生存期的关系。两组间比较采用Mann-Whitney U检验,四组间比较采用Kruskal-Wallis检验。结果自建的点免疫印迹法检测血清外泌体ANXA11具有较高的敏感度和重复性。利用构建的方法检测研究对象发现ANXA11在PC组升高最明显,与其余三组比较,差异有统计学意义(Hc=58.079,P<0.01)。ROC曲线表明,ANXA11(AUC=0.836)对PC的诊断效能高于CEA(AUC=0.656),与CA19-9(AUC=0.870)相当,联合CA19-9可提高诊断PC的敏感度,且保持较高的特异度。PC患者治疗前血清外泌体ANXA11水平与年龄、性别、肿瘤大小、肿瘤生长部位、淋巴结转移、远处转移及TNM分期均无关(Z值分别为0.052、-0.285、-0.402、0.324、0.888、0.658、1.734,P均>0.05),且PC患者术后第10天血清外泌体ANXA11水平与术前相比,差异无统计学意义(Z=-1.569,P=0.12)。PC患者生存时间与是否存在淋巴结转移、远处转移、TNM分期及治疗方式有关(χ^2分别为9.354、6.086、9.389、16.998,P均<0.05),而与ANXA11、CA19-9及CEA等无相关性(χ^2分别为1.516、0.011、0.159,P均>0.05)。结论本研究成功建立了检测人血清外泌体ANXA11的方法。血清外泌体ANXA11联合CA19
Objective To explore a method for detecting recombinant human Annexin A11(ANXA11)in serum exosomes of pancreatic cancer patients,and then primarily evaluate the clinical value of ANXA11 in pancreatic cancer patients.Methods A prospective study was conducted and serum specimens from 70 patients diagnosed with PC,15 patients diagnosed with benign pancreatic mass and 70 patients diagnosed with pancreatitis from the Affiliated Hospital of Nantong University were collected from December 2016 to July 2019.70 healthy subjects during the same period were selected as control group.The abundance of ANXA11 in serum and exosomes-free serum were detected through parallel reaction monitoring(PRM)basing on high-resolution,high-precision mass spectrometer.Dot immunoblotting created by ourselves for detecting ANXA11 in exosomes and then the methodological evaluation were carried out.Levels of ANXA11 in exosomes in all subjects were statistically analyzed.Moreover,the areas under the curve(AUC)of receiver operating characteristic(ROC)curves were adopted to evaluate the diagnostic efficacy of ANXA11,CA19-9,CEA on PC.The relationship between ANXA11 and clinicopathological parameters as well as prognosis of PC patients was analyzed in the next moment.For analysis,the Mann-Whitney U test was used for comparing between either two groups,and the kruskal-wallis test was used for comparison among four groups.Results The detection of serum exosome ANXA11 has high sensitivity and repeatability by the method of self-established dot immunoblotting.ANXA11 increased most significantly in the PC group,and the difference was statistically significant(Hc=58.079,P<0.01)compared with other three groups.ROC curve analysis showed that the diagnostic performance of ANXA11 area under the curve(AUC=0.836)was higher than CEA(AUC=0.656)and equal to CA19-9(AUC=0.870).The combination of ANXA11 and CA19-9 could improve the sensitivity of diagnosing PC and maintain good specificity.The level of serum exosome ANXA11 before treatment in PC patients was not relat
作者
谢旭东
徐伟松
陈晓君
刘肇修
徐晓耘
鲍柏军
丛辉
肖明兵
Xie Xudong;Xu Weisong;Chen Xiaojun;Liu Zhaoxiu;Xu Xiaoyun;Bao Baijun;Cong Hui;Xiao Mingbing(Department of Gastroenterology,Affiliated Hospital of Nantong University,Nantong 226001,China;Department of Gastroenterology,Nantong Second People′s Hospital,Nantong 226002,China;Office of Infection Management,Affiliated Hospital of Nantong University,Nantong 226001,China;Department of Medical Laboratory,Affiliated Hospital of Nantong University,Nantong 226001,China)
出处
《中华检验医学杂志》
CAS
CSCD
北大核心
2020年第6期659-665,共7页
Chinese Journal of Laboratory Medicine
基金
江苏省科技重点研发面上项目(BE2019692)
中国博士后基金项目(2019M661909)
江苏省卫健委面上项目(H2019072)
南通市科技计划项目(MS22018006,MS12019018,MS12019020,MSZ19177)。
关键词
膜联蛋白A11
胰腺癌
血清外泌体
诊断
肿瘤标志物
Recombinant human Annexin A11
Pancreatic cancer
Serum exosomes
Diagnosis
Tumor markers
