外周血循环肿瘤细胞和游离DNA联合检测在乳腺癌辅助诊断中的临床价值  

Clinical diagnostic value of circulating tumor cells and circulating cell-free DNA combined detection in peripheral blood for breast cancer

作　　者：张潇分 贾春平[2] 陈宏梅[3] 施英娟[1] 鞠少卿[1] 王旭东[1] 储海丹[1] 丛辉[1] Zhang Xiaofen;Jia Chunping;Chen Hongmei;Shi Yingjuan;Ju Shaoqing;Wang Xudong;Chu Haidan;Cong Hui(Department of Laboratory Medicine,Affiliated Hospital of Nantong University,Nantong 226001,China;Shanghai Institute of Microsystem and Information Technology,Chinese Academy of Sciences,Shanghai 200050,China;Department of Special Medical Treatment,VIP ward,Affiliated Hospital of Nantong University,Nantong 226001,China)

机构地区：[1]南通大学附属医院医学检验科,南通226001 [2]中国科学院上海微系统与信息技术研究所,上海200050 [3]南通大学附属医院特需病区,南通226001

出　　处：《中华检验医学杂志》2019年第8期662-668,共7页Chinese Journal of Laboratory Medicine

基　　金：江苏省医学重点学科(ZDXKB2016011);江苏省六大人才高峰项目(WS066);南通市科技项目(MS12017008-1).

摘　　要：目的探讨外周血液中循环肿瘤细胞(CTCs)和循环游离DNA(cfDNA)检测在乳腺癌患者中的临床价值.方法收集2017年7月至2018年4月就诊于南通大学附属医院的47例乳腺癌患者(Ⅱ期7例、Ⅲ期19例、Ⅳ期21例),24例乳腺良性疾病患者,以及同期28名健康体检者外周血液标本.采用基于尺寸的高通量微流控芯片捕获CTCs、基于Alu序列的实时荧光定量PCR检测游离DNA长、短片段(247bp、115bp),并以长、短片段扩增产物比值作为DNA完整性指标.采用Mann-WhitneyU检验或Kruskal-WallisH检验比较各组差异,分析CTCs和cfDNA与乳腺癌临床参数的关系.绘制ROC曲线并计算曲线下面积(AUC)用于评价血细胞CTCs及血浆cfDNA检测作为诊断标准的可行性.结果47例乳腺癌患者CTCs(13.98±12.36)个/ml、cfDNA的完整性(Alu247/115)(0.7687±0.3868)与正常对照组的(1.14±1.35)个/ml、0.5094±0.2456相比,差异有统计学意义(U值分别为126.5、359.0,P均<0.001);ROC曲线下面积为:CTCs:0.885(95%CI:0.805~0.965),cut-off值为7.68个/ml,灵敏度为80.4%,特异度为96.4%;Alu247/115:0.727(95%CI:0.608~0.847),cut-off值为0.431,灵敏度为71.7%,特异度为71.4%.CTCs和Alu247/115联合检测时曲线下面积为0.919(95%CI0.854~0.984),高于各指标单项检测.结论CTCs与cfDNA可能是乳腺癌辅助诊断的潜在生物学指标,CTCs与cfDNA联合检测可能提高乳腺癌患者的诊断率.Objective To investigate the clinical diagnostic value of circulating tumor cells (CTCs) and circulating cell-free DNA (cfDNA) in peripheral blood samples in breast cancer. Methods From July 2017 to April 2018, 47 patients with BMC (7 in stage Ⅱ, 19 in stage Ⅲ and 21 in stage Ⅳ), 24 patients with benign breast diseases and 28 healthy people were selected. After collecting peripheral blood samples, serum and blood cells were separated. The size-based high-throughput microfluidic chip was used to capture CTCs. The real-time fluorescent quantitative PCR based on Alu sequence was used to detect the length of cfDNA (247 bp, 115 bp) in the serum, and the ratio of amplified products of long and short fragments was used as the index of DNA integrity. The Mann-Whitney U test or Kruskal-Wallis H test was used to compare the differences between the groups and analyze the relationship between CTCs and cfDNA and clinical parameters of breast cancer. The ROC curve was drawn and the area under the curve (AUC) was used to evaluate the feasibility of blood cell CTCs and plasma cfDNA detection as diagnostic criteria. Results The CTCs and cfDNA of 47 BMC patients were analyzed. The CTCs and cfDNA integrity index (Alu 247/115) of BMC patients were significantly higher than those of physical examination patients[(13.98±12.36)cells/ml vs (1.14±1.35) cells/ml;0.768 7±0.386 8 vs 0.509 4±0.245 6], and the difference was statistically significant(the U value was 126.5, 359.0;P<0.001), the area under ROC curve of CTCs was 0.885 (95%CI: 0.805-0.965), cut-off value was 7.68/ml, sensitivity was 80.4%, specificity was 96.4%. The area under ROC curve of Alu 247/115 was 0.727(95%CI: 0.608-0.847), cut-off value was 0.431, sensitivity was 71.7%, specificity was 71.4%. The AUC of CTCs and Alu 247/115 was 0.919 (95%CI 0.854-0.984), which was higher than the single test of each indicator. Conclusions CTCs and cfDNA may be the potential biological indicators for breast cancer diagnosis. The combined detection of CTCs and cfDNA maybe improve

关 键 词：乳腺肿瘤 肿瘤细胞 循环 游离核酸 生物标记 肿瘤 

分 类 号：R737.9[医药卫生—肿瘤] R730.43[医药卫生—临床医学]