乳腺癌循环肿瘤细胞

循环肿瘤细胞的播散是乳腺癌转移与复发的重要事件.检测循环肿瘤细胞,对判断患者预后、评价治疗效果、监测转移复发、指导治疗等有着重要的临床意义.完善循环肿瘤细胞的检测技术,研究循环肿瘤细胞的生物学特性及其机制,是乳腺癌循环肿瘤细胞研究需要解决的主要难题.目前,乳腺癌循环肿瘤细胞研究在检测技术及其指标的联合应用、低增殖活性循环肿瘤细胞研究等方面均取得了一定进展.     

循环肿瘤细胞及其与肿瘤转移复发的相关性研究

肿瘤细胞血液播散至远处器官和后来形成的转移灶是上皮来源恶性肿瘤最主要的死亡原因。对恶性肿瘤发生远处转移和复发等研究认为是由外周血中存在的循环肿瘤细胞所致(circulating tumor cell,CTC)。CTCs存在于外周循环中,激发机体的免疫反应,大部分肿瘤细胞被机体免疫系统所识别难以生存,只有小部分细胞继续在循环中,穿透基底膜,侵出血管在远端形成肿瘤转移灶。CTCs是肿瘤发生远处转移的必要条件,深入研究循环肿瘤细胞有助于对肿瘤的转移机制有更多的了解。本文对CTCs的分离与鉴定技术方法和CTCs与肿瘤复发转移的机制进行综述。     

循环肿瘤细胞检测与结直肠癌复发转移的研究进展

结直肠癌发病率在逐年上升,转移和复发是结直肠癌死亡的主要原因。血液循环中的肿瘤细胞可能具有增生能力或转移潜能,循环肿瘤细胞（circulating tumor cell,CTC）的检测为预测评估复发转移及疗效、肿瘤分期及个体化治疗提供了一个可靠手段,随着检测技术的不断优化发展,将为临床提供更好的支持。本文旨在对CTC检测与结直肠癌复发转移等方面的研究进展进行综述。      

循环肿瘤细胞及其与肿瘤转移复发的相关性研究

肿瘤细胞血液播散至远处器官和后来形成的转移灶是上皮来源恶性肿瘤最主要的死亡原因。对恶性肿瘤发生远处转移和复发等研究认为是由外周血中存在的循环肿瘤细胞所致（circulating tumor cell,CTC）。CTCs存在于外周循环中,激发机体的免疫反应,大部分肿瘤细胞被机体免疫系统所识别难以生存,只有小部分细胞继续在循环中,穿透基底膜,侵出血管在远端形成肿瘤转移灶。CTCs是肿瘤发生远处转移的必要条件,深入研究循环肿瘤细胞有助于对肿瘤的转移机制有更多的了解。本文对CTCs的分离与鉴定技术方法和CTCs与肿瘤复发转移的机制进行综述。     

循环肿瘤细胞检测及临床应用的研究进展

研究发现，实体瘤患者外周血中存在循环肿瘤细胞。循环肿瘤细胞的检测有助于发现早期肿瘤患者的微转移，重新确定临床分期，监测患者术后肿瘤是否复发与转移，评估预后，选择个体化的治疗策略。由于存在循环肿瘤细胞检测方法、原发肿瘤细胞的不连续脱落、基因组的不稳定性及无效转移等诸多问题，循环肿瘤细胞检测的生物学意义和临床意义仍存在很大争议。本文综述了对实体瘤患者循环肿瘤细胞检测的方法和结果，并探讨当前肿瘤研究领域面临的问题和潜在的进展。     

检测循环肿瘤细胞(CTC)在临床肿瘤诊断和治疗中的作用

有关循环肿瘤细胞的理论已有很长时间。肿瘤转移及复发的过程均是一个循环肿瘤细胞定植生长的级连放大的瀑布反应。目前肿瘤的发现和诊断仍高度依赖于影像学及肿瘤特异性血清标志物的方法,不能及时发现肿瘤的转移和复发。基于肿瘤细胞播散理论,检测循环肿瘤细胞可更早期有效发现肿瘤的早期转移和复发。本文对文献报道的检测循环肿瘤细胞在临床肿瘤诊断和治疗中的作用进行了总结,并与目前经典的标志物和影像学进行了比较。     

循环肿瘤细胞簇在肿瘤转移中的研究进展

肿瘤转移是肿瘤相关死亡的主要原因,大量研究显示循环肿瘤细胞（circulating tumor cells,CTCs)在肿瘤转移中起着重要作用。越来越多的研究表明循环肿瘤细胞簇（circulating tumor cells clusters,CTC clusters）与单循环肿瘤细胞相比转移能力更强。分离检测技术的不断进步为深入研究循环肿瘤细胞簇提供了便利。本文就循环肿瘤细胞簇在肿瘤转移中的研究进展作一综述。     

乳腺癌循环肿瘤细胞检测及其研究进展

循环肿瘤细胞(CTC)为肿瘤发生转移的重要原因.检测CTC能早期发现肿瘤的转移及复发.在乳腺癌中,CTC检测与其他检测方式相比有较高的可重复性和敏感性,并能更早地提供预后信息.     

结直肠癌循环肿瘤细胞及DNA的研究进展

结直肠癌是最常见的恶性肿瘤之一,尽管联合应用手术、化疗、放疗可提高患者的生存率,但仍有大量患者因术后转移和复发死亡。循环肿瘤细胞及循环肿瘤DNA对结直肠癌的早期诊断、疗效监测、预后评估以及个体化治疗方案制定均有重要意义,已成为近年来研究的热点。本文对近年来结直肠癌循环肿瘤细胞及循环肿瘤DNA的检测技术及其在早期诊断、疗效监测和预后判断中的应用进展作一综述。     

循环肿瘤细胞和循环肿瘤内皮细胞计数对局限性肾癌术后复发转移监测的价值

目的:探讨局限性肾癌患者行根治或部分切除术后循环肿瘤细胞和循环肿瘤内皮细胞数量变化在预测肿瘤复发转移中的作用。方法:收集52例局限性肾癌患者术前和术后3个月7.5 mL外周血和20例健康者7.5 mL外周血。术后循环肿瘤细胞数量和循环肿瘤内皮细胞数量高于术前者纳入阳性组,反之纳入阴性组。分别对两组患者进行随访,终点事件为肿瘤复发转移,最长随访时间21个月。结果:CTC阳性组复发转移率为61.9%（13/21）,明显高于阴性组19.4%（6/31）,差异有统计学意义(P=0.003),CTEC阳性组复发转移率为57.1%（12／21）,阴性组复发转移率为22.6%（7/31）,差异有统计学意义（P=0.012）,双阳性组复发转移率为75.0%(12/16)。结论:术后3个月循环肿瘤细胞计数高于术前的肾癌患者易出现复发转移,若同时术后循环肿瘤内皮细胞计数高于术前,则提示患者复发转移率可能会更高。循环肿瘤细胞和循环肿瘤内皮细胞数量变化可作为较好的肾癌术后监测指标。     

高灵敏度hMAM-QDs脂质体结合CK19双标记法检测乳腺癌循环肿瘤细胞的临床研究

目的:评估细胞角蛋白19（CK19）单标记法和CK19结合人乳球蛋白-量子点（hMAM-QDs）双标记法检测乳腺癌循环肿瘤细胞（circulating tumor cells,CTCs）的临床价值。方法:纳米脂质体（PL）技术用于构建hMAM-PL（QDs）;借助免疫磁珠分离技术,使用上皮细胞黏附分子（EpCAM）抗体分离乳腺癌CTCs。结果:hMAM-PL（QDs）［平均粒径（102.6±1.1）nm,多分散指数0.047］可以显著提高实验中的检测灵敏度。此外,CK19-FITC与hMAM-PL（QDs）双标记方法相结合可有效区分分离的CTCs,提高准确性,并减少乳腺癌检测中的假阳性。CTCs编号、图像与临床特征以及病理状况高度一致。结论:CK19结合hMAM-PL（QDs）双标记方法检测比CK19单标记方法更准确。双标记方法可以显著减少假阳性结果,对于早期乳腺癌的诊断、治疗和预后具有重要意义。     

结直肠癌患者循环肿瘤细胞与临床病理特征的关系

目的:探讨外周血循环肿瘤细胞（circulating tumor cells,CTCs）在结直肠癌患者检测中的临床意义。方法:收集陕西省人民医院结直肠癌患者外周血标本57例,良性病变和健康受试者外周血标本29例。应用CyttelTM-CTC平台检测技术（即阴性富集技术结合免疫荧光原位杂交法）检测受试者外周血CTCs,搜集结直肠癌患者临床病理特征,采用统计学方法分析与其外周血CTCs之间的关系。结果:在Cut-off值≥2的结直肠癌患者中CTCs检出率为78.9%（45/57）,远高于良性病变及健康受试者中CTCs检出率的6.9%（2/29）（P＜0.05）。在Cut-off值≥3的结直肠癌患者中,发生淋巴转移的患者CTCs检出率为62.1%（18/29）,高于未发生淋巴转移患者CTCs检出率的32.1%（9/28）（P=0.024）。在Cut-off值≥4的结直肠癌患者中,周围组织侵犯患者CTCs检出率为53.8%（7/13）,高于周围组织未侵犯患者CTCs检出率的25.0%（11/44）（P=0.049）。在不同的病理分期中,与癌胚抗原（CEA）检测比较,CTCs检出率均高于CEA检出率。结论:CTC平台检测技术可有效检测结直肠癌患者外周血CTCs,CTCs计数分布与结直肠癌患者的病理分期显著相关,该技术敏感性高,准确性好,对于结直肠癌患者的临床诊断具有重要指导意义。     

Morphological Features of Breast Cancer Circulating Tumor Cells in Blood After Physical and Biological Type of Isolation

BackgroundCirculating tumor cells (CTCs) have become an important biomarker in breast cancer. Different isolation tech-niques based on their biological or physical features were established. Currently, the most widely used methods for visualization after their separation are based on immunofluorescent staining, which does not provide the information on the morphology.Materials and methodsThe aim of this study was to evaluate how two different separation techniques affect cell morphology and to analyse cell morphology with techniques used in routine cytopathological laboratory. A direct side-by-side comparison of physical (Parsortix®) and biological (MACS®) separation technique was performed.ResultsIn the preclinical setting, both isolation techniques retained the viability and antigenic characteristics of MCF7 breast cancer cells. Some signs of degeneration such as cell swelling, cytoplasmic blebs, villous projections and vacuolization were observed. In metastatic breast cancer patient cohort, morphological features of isolated CTCs were dependent on the separation technique. After physical separation, CTCs with preserved cell morphology were detected. After biological separation the majority of the isolated CTCs were so degenerated that their identity was difficult to confirm.ConclusionsTaken together, physical separation is a suitable technique for detection of CTCs with preserved cell morphology for the use in a routine cytopathological laboratory.Key words: CTC, breast cancer, morphology, Parsortix®, MACS®, Giemsa     

循环肿瘤细胞的富集分离和检测

循环肿瘤细胞（circulating tumor cells,CTCs）与恶性肿瘤的复发和转移有密切关系。CTCs 是指自发或因诊疗操作由实体瘤或转移灶释放进入外周血循环的肿瘤细胞。但是,CTCs 在癌症患者外周血中含量极少而且其在血液循环中往往相互聚集或者与其他细胞黏附集聚,另外循环肿瘤细胞还可能发生一些转变。例如上皮－间质转化（epithelial －mesenchymal transition,EMT）,造成的 CTCs 表面标志物的改变,因此CTCs 的富集分离、检测十分困难。诸多技术各有各的优缺点,至今还没有统一的标准。本文就当今正使用的一些富集分离和检测技术进行总结并分析其优缺点。     

肺癌循环肿瘤细胞的研究进展

转移是导致肺癌患者死亡的主要因素,积极防治转移的发生是提高患者疗效的关键。循环肿瘤细胞（CTCs）是启动远处转移的重要环节,自原发灶脱落开始即触发一系列转移级联反应,最终在远处靶器官定植和增殖。近年来,肺癌CTCs的检测技术手段不断优化,不仅可以对CTCs进行计数,还可鉴定不同亚群。肺癌CTCs检测可用于早期筛查、手术及放化疗后的预后预测、免疫靶向治疗的疗效评价等方面,动态监测肿瘤异质性,有助于调整个体化治疗方案,达到精准治癌的目的。田建辉研究组建立世界首例人肺腺癌循环肿瘤细胞系,将其融入肺癌亚临床核心病机“正虚伏毒”学说的探索,并积极促进肺癌特异性研究平台的构建,以期提高肺癌转移的研究和防控效率。     

Clinical implications and perspectives of portal venous circulating tumor cells in pancreatic cancer

Despite recent improvements in the diagnosis and treatment of pancreatic cancer(PC), clinical outcomes remain dismal. Moreover, there are no effective prognostic or predictive biomarkers or options beyond carbohydrate antigen 19-9 for personalized and precise treatment. Circulating tumor cells(CTCs), as a member of the liquid biopsy family, could be a promising biomarker; however, the rarity of CTCs in peripheral venous blood limits their clinical use. Because the first venous drainage of PC is ...     

Separation of circulating cancer cells by unique microfluidic chip in colorectal cancer

Circulating tumor cells (CTCs) from peripheral blood are emerging as a useful tool for the detection of malignancy, monitoring disease progression, and measuring response to therapy. We describe a unique microfluidic chip that was capable of efficient and selective separation of CTCs from peripheral whole blood samples. The ability of microfluidic chip to capture CTCs from PBS and whole blood samples was tested. Sixty-eight peripheral blood samples from 68 colorectal cancer patients were investigated for the presence of CTCs by microchip technology. The frequency of CTCs was analyzed statistically for correlation with relevant clinical data. We also examined samples from 20 healthy individuals as controls. The calculated capture efficiency was 85.7% and decreased significantly at flow rates above 2.0 ml/h. The number of CTCs isolated ranged from 3 to 236/ml for colorectal patients [99 +/- 64 (mean +/- SD) CTCs/ml]. None of the 20 healthy subjects had any identifiable CTCs. We identified CTCs in 46 (67.65%) of the 68 patients: in two of nine (22.22%) Dukes A, in 10 of 24 (41.67%) Dukes B, in 21 of 22 (95.45%) Dukes C, and in all 13 Dukes D patients. The detection rate in Dukes C and D patients was much higher than in Dukes A and B patients (97.73% vs. 36.36%) (p < 0.01). A significant correlation between detection of CTCs and clinical stage (r = 0.792, p < 0.01) was found, which was higher than carcinoembryonic antigen (r = 0.285, p > 0.01), carbohydrate antigen 19-9 (r = 0.258, p > 0.01), alpha-fetoprotein (r = 0.096, p > 0.01), and cancer antigen 125 (r = 0.134, p > 0.01). Microfluidic chip provides a novel method for capturing CTCs. The presence of CTCs correlated with clinical stage. It is important to evaluate CK-positive and DAPI-stained tumor cells together to determine the role of CTCs in tumor behavior and disease progression.     

Role of Circulating Tumor Cells in Metastatic Colorectal Cancer: Clinical Challenges and Opportunities

Circulating tumour cells (CTCs) have been presumed critical to the metastatic process since 1800. These epithelial cells are disseminated from the primary or metastatic tumor site and can be identified in the peripheral blood of patients. Nowadays, technical advances have rendered it easier to reproducibly and repeatedly sample this population of cells with an acceptable degree of accuracy. Therefore, these cells could become an attractive surrogate for phenotypic and genotypic markers in correlation with the development of molecular targeted therapies. In metastatic colorectal cancer several prospective studies have demonstrated the independent prognostic significance of CTCs and identified the number of CTCs before and during treatment as a predictor for overall survival and disease free survival. However, the underlying molecular characteristics of CTCs associated with outcome remain largely unknown. In this review, the role of CTCs in metastatic colorectal cancer is discussed. The variety of assays that can be used for enrichment and detection steps in CTC detection will be described and the clinical utility of these cells for assessing prognosis and monitoring response to therapy will be analyzed. We will also address the shortcomings of current detection methods that fail to identify a mesenchymal subgroup of CTCs, and briefly address how characterization of these cells can help elucidate the biology of cancer metastases.     

循环肿瘤细胞检测在结直肠癌临床中的应用

结直肠癌是常见的恶性肿瘤之一,其复发和转移是导致患者死亡的主要原因。外周血循环肿瘤细胞（circulating tumor cells,CTCs）是导致结直肠癌患者复发转移的关键性因素之一,在结直肠癌患者复发转移的监测、预后、病情评估以及治疗等临床问题中有重要的应用价值,已成为近年来研究的热点。本文从CTCs的分离富集、检测技术及在结直肠癌患者中的临床应用价值等几个方面,全面阐述了CTCs检测在结直肠癌临床应用中的研究进展。