循环肿瘤细胞的检测及其在乳腺癌患者中的临床应用

由于循环肿瘤细胞（ｃｉｒｃｕｌａｔｉｎｇｔｕｍｏｒｃｅｌｌ,ＣＴＣ）能够反映肿瘤侵袭与转移,检测ＣＴＣ也就在成为肿瘤界研究的热点。外周血ＣＴＣ非常少,仅能通过肿瘤特异性抗原来检测。本综述主要介绍了ＣＴＣ的富集、检测过程、常用标记物及其在乳腺癌患者中的临床应用。     

Evaluation of circulating tumor cells in patients with breast cancer: multi-institutional clinical trial in Japan

BACKGROUND: With the development of the CellSearch System, it has become possible to measure circulating tumor cell (CTC) levels with high reproducibility, and the CTC test is currently being used clinically for patients with metastatic breast cancer in the United States. It is imperative that the clinical significance of the CTC test also be examined in Japan. METHODS: Using the CellSearch System, CTC levels were evaluated in 57 healthy individuals and patients with benign breast disease; 30 patients with primary breast cancer (stages 1-3); and 38 patients with metastatic breast cancer. First, the relationship between CTC levels and the presence of metastasis was examined using a cutoff score of 2 CTCs per 7.5 ml whole blood. Then, the patients with metastatic breast cancer were divided into two groups, using a cutoff score of 5 CTCs per 7.5 ml blood, and progression-free survival (PFS) and overall survival (OS) were compared in the two groups. RESULTS: When the clinical cutoff score was set at 2 CTCs per 7.5 ml blood, 0% of the healthy individuals and patients with benign breast disease (0/57), 3.3% of the patients with primary breast cancer (1/30), and 50% of the patients with metastatic breast cancer (19/38) were identified as as having 2 CTCs per 7.5 ml blood. Additionally, with a cutoff score of 5 CTCs, 11 patients were reported to have 5 or more CTCs and both PFS (P = 0.0036) and OS (P = 0.04) were worse for this patient population than for the population with fewer than 5 CTCs. CONCLUSION: As concluded in a similar clinical trial in the United States, for patients with breast cancer, measuring CTC levels can be both an accurate indicator of metastases and an important measure of patient prognosis.     

HER2 status predicts the presence of circulating tumor cells in patients with operable breast cancer

Introduction Circulating tumor cells (CTCs) correlate with worse prognosis in patients with metastatic breast cancer, but there are little data on CTCs in operable patients. We hypothesized that primary tumor characteristics would predict the likelihood of identifying CTCs in patients with operable breast cancer. Methods Clinical and pathological data from 92 patients with operable breast cancer were collected. The CellSearch system was used to detect CTCs in 30 ml of peripheral blood. CTCs were defined as nucleated cells lacking CD45 but expressing cytokeratins 8, 18, or 19. Univariate analysis was performed to determine if the presence of any primary tumor characteristic was predictive of CTCs. As a secondary objective we evaluated if nodal or bone marrow status was predictive of CTCs. Results Thirty-eight percent of patients (35/92) had evidence of CTCs, with a median number of 1.0 CTC per CTC positive patient (range 1–22). HER2 status was the sole primary tumor characteristic that reliably predicted the presence of CTCs (P = 0.01, risk ratio = 3.66). No significant association was found between the presence of CTCs and tumor size (T), estrogen receptor (ER) status, progesterone receptor (PR) status, grade, histologic type, degree of nodal involvement (N), lymphovascular invasion (LVI) or Ki-67 proliferation. Bone marrow micrometastases were found in 17/64 (26.6%) of the patients but did not correlate with the presence of CTCs. Conclusion HER2 status was the only primary tumor characteristic that correlated with the presence of CTCs. Long-term follow-up will be required to determine the significance of CTCs in operable breast cancer. Presented in part at the 2007 Society of Surgical Oncology Cancer Symposium.     

Circulating tumor cells in breast cancer: methodology and clinical repercussions

Breast cancer is the most common type of cancer among women, and clinicians have long recognized its heterogeneity. Its detection and treatment in early stages allow for reduction of mortality. Despite the advances and new strategies for combining surgical, radiotherapy, and chemotherapy options, however, the percentage of patients developing metastases and advanced stages remains high. Even though serum tumor markers have been used for the early diagnosis of metastases, their systematic determination has not had an effect on survival. Methods that are more reliable are needed to detect metastases earlier than with the common clinical methods and thus start treatment before overt relapse. Early indicators of response or resistance to treatment are also an issue in clinical practice. Imaging techniques are time consuming, and it is difficult to detect changes that indicate response limited to therapy, and approaches to defining changes in tumor mass are time and resource consuming. In contrast, detection of circulating tumor cells (CTC) could be a useful tool in early detection of relapse and response to systemic chemotherapy. Extremely sensitive techniques are available that are easily applied to peripheral blood samples, which might provide enormous research possibilities in this area.     

CD49f-based selection of circulating tumor cells (CTCs) improves detection across breast cancer subtypes

Circulating tumor cells (CTCs) can be enumerated using CellSearch, but not all breast cancer subtypes, specifically those with epithelial-mesenchymal transition (EMT) characteristics, sufficiently express the enrichment (EpCAM) and selection (CK8/18/19) markers used in this method. While CD146 can detect EpCAM-negative CTCs, we here evaluated the value of various cytokeratins and CD49f to detect CK8/18/19-negative CTCs. The tested cytokeratins provided no substantial benefit, but adding CD49f to CK8/18/19 as a selection marker resulted in improved recovery of normal-like cell lines.     

Monitoring apoptosis and Bcl-2 on circulating tumor cells in patients with metastatic breast cancer

BackgroundEnumeration of circulating tumor cells (CTC) from whole blood permits monitoring of patients with breast carcinoma. Analysis of apoptosis & Bcl-2 expression in CTC might add additional prognostic and predictive information. We estimated the degree of these markers in CTC from patients being treated for metastatic breast cancer.MethodsEighty-three evaluable patients initiating a new therapy for metastatic breast cancer were enrolled. Whole blood was collected at baseline, at one of three short term time windows (24, 48, or 72 h) after initiating treatment, and at first follow-up (3–5 weeks). CTC were isolated, enumerated, and expression of M30 and Bcl2 was determined using the CellSearch^® System.ResultsAt baseline, window, and 3–5 weeks post-treatment, 41/80 (51%), 40/80 (50%) and 21/75 (28%) patients had ≥5 CTC, respectively. At baseline, the proportion of CTC-apoptosis (M30) was inversely correlated with CTC number, and modestly inversely correlated with CTC-Bcl-2. As expected, higher CTC levels at baseline or first follow-up were associated with worse prognosis. Surprisingly, in patients with elevated CTC, higher levels of CTC-apoptosis were associated with worse prognosis, while higher CTC-Bcl-2 levels correlated with better outcomes.ConclusionsCTC apoptosis and expression of Bcl-2 can be analytically determined in patients with metastatic breast cancer and may have biological and clinical implications. Characterization of CTC for these and other markers could further increase the utility of CTC monitoring patients in clinical investigations of new anti-neoplastic agents.     

循环肿瘤DNA在晚期乳腺癌的临床应用进展

晚期乳腺癌是严重威胁全世界女性健康的第一大恶性肿瘤,目前以综合治疗为主,包括化疗、内分泌治疗和靶向治疗等。短期内复发、转移是限制临床疗效和导致患者预后差的重要因素,而影像学检查和传统血清标记物却难以及时有效地监测肿瘤的复发转移,导致临床治疗决策迟滞。因此,研发灵敏、准确、非／低侵入性的疗效监测方法 至关重要。循环肿瘤DNA（ctDNA）是游离于血浆中的来源于肿瘤细胞的坏死、凋亡和分泌过程的DNA。多项研究证实检测ctDNA突变水平变化对于恶性肿瘤早期诊断、用药指导、复发监测有着巨大的应用潜力。本文将对ctDNA监测晚期乳腺癌的临床应用研究进展综述如下。     

循环肿瘤细胞在转移性乳腺癌中的表达及其与多种血清肿瘤标志物的关系

目的 探讨转移性乳腺癌的循环肿瘤细胞(CTC)和血清肿瘤标志物(CEA、CA153和CA125)的表达水平以及两者之间的相关性。方法 采用CellSearch自动检测系统检测93例转移性乳腺癌患者开始新治疗前的循环肿瘤细胞(CTC),同时采用电化学发光法检测这些患者血清肿瘤标志物(CEA、CA153和CA125)的表达水平。结果 CTC阳性率为60%(56/93),与患者激素受体的状态、Her-2的状态、既往治疗的线数以及内脏转移无关。CEA的阳性率为56%(52/93),ER或PR阳性的患者CEA的阳性率更高(χ²=4.550,P=0.045),与Her-2的状态、既往治疗的线数以及内脏转移无关;CA153的阳性率为47%(44/93),与患者激素受体的状态、Her-2的状态、既往治疗的线数以及内脏转移无关;CA125的阳性率为41%(38/93);既往治疗大于等于二线的患者CA125的阳性率更高(χ²=4.501,P=0.038),与患者激素受体的状态、Her-2的状态以及是否有内脏转移无关。CTC和CEA之间呈正相关(r=0.296,P=0.004);CTC和CA153之间呈正相关(r=0.286,P=0.005);CTCs和CA125两者之间无相关性(r=0.184,P=0.077)。结论 转移性乳腺癌患者的循环肿瘤细胞(CTC)检出率高,而且与血清肿瘤标志物(CEA和CA153)有明显相关性,提示联合检测转移性乳腺癌的循环肿瘤细胞(CTC)和血清肿瘤标志物(CEA和CA153)可能会对其治疗决策有所帮助,同时CTC有可能成为转移性乳腺癌新的治疗靶点。     

液体肿瘤生物指标检测对乳腺癌患者的临床价值

循环肿瘤标志物在乳腺癌中的应用受到越来越多的关注,研究水平已从蛋白质水平深入到基因水平,相应的检测指标则从传统的肿瘤标志物扩展到相对特异的HER-2蛋白质细胞外段、循环肿瘤细胞、循环肿瘤DNA及循环RNA等。作为“液体检测”,循环肿瘤标志物的检测因其实时动态、操作简便、可重复性好等优势,被广泛应用于协助早期诊断、判断预后、预测疗效、监测疾病复发及病情变化等方面。对循环肿瘤标志物的深入研究,有助于实现患者的个体化治疗。     

循环肿瘤DNA检测在乳腺癌诊断中的临床应用

随着二代测序技术的发展,循环肿瘤DNA(circulating tumor DNA,ctDNA)检测在乳腺癌中的应用得到越来越多的关注。目前国内外大量研究显示ctDNA检测技术在监测肿瘤负荷、疗效预测、早期诊断、预后分析等方面具有广阔的应用价值。在乳腺癌诊疗走向个体化精准的时代,ctDNA检测能够为患者提供更为精准的诊断,指导临床治疗。     

The clinical significance of circulating tumor cells in non-metastatic colorectal cancer - A review

Background

Finding a clinical tool to improve the risk stratification and identifying those colorectal cancer patients with an increased risk of recurrence is of great importance. The presence of circulating tumor cells (CTC) in peripheral blood can be a strong marker of poor prognosis in patients with metastatic disease, but the prognostic role of CTC in non-metastatic colorectal cancer is less clear. The aim of this review is to examine the possible clinical significance of circulating tumor cells in non-metastatic colorectal cancer (TNM-stage I-III) with the primary focus on detection methods and prognosis.

Methods

The PubMed and Cochrane database and reference lists of relevant articles were searched for scientific literature published in English from January 2000 to June 2010. We included studies with non-metastatic colorectal cancer (TNM-stage I-III) and CTC detected pre- and/or post-operatively in peripheral blood.

Results

Nine studies qualified for further analyses. Detection rates of CTC in peripheral blood of patients with non-metastatic colorectal cancer varied from 4% to 57%. Seven studies applied RT-PCR and two studies used immunocytochemical methods. Seven studies found the presence of CTC to be a prognostic marker of poor disease-free survival.

Conclusion

The presence of CTC in peripheral blood is a potential marker of poor disease-free survival in patients with non-metastastic colorectal cancer. The low abundance of CTC in non-metastatic colorectal cancer requires very sensitive and specific detection methods. An international consensus on choice of detection method and markers, is warranted before incorporating CTC into risk stratification in the clinical setting.     

Circulating tumor cells with a putative stem cell phenotype in peripheral blood of patients with breast cancer

The CD44⁺/CD24^−/low and ALDH1⁺ cell phenotypes are associated with stemness and enhanced tumorigenic potential in breast cancer. We assessed the expression of CD44, CD24 and ALDH1 on tumor cells circulating in the peripheral blood (CTCs) of patients with metastatic breast cancer using triple-marker immunofluorescence microscopy. Among a total of 1439 CTCs identified in 20 (66.7%) out of 30 patients, 35.2% had the stem-like/tumorigenic phenotype CD44⁺/CD24^−/low, whereas 17.7% of the CTCs analyzed in seven patients, were ALDH1^high/CD24^−/low. In conclusion, we report the existence of a subpopulation of CTCs with putative stem cell progenitor phenotypes in patients with metastatic breast cancer.     

循环肿瘤细胞在肺癌中的研究

循环肿瘤细胞(CTC)是存在于外周血中的肿瘤细胞,在肺癌复发及转移过程中有重要作用.随着目前对CTC的研究及靶向治疗的不断深入,发现CTC可作为一种“液态”肿瘤组织指导临床治疗,对肺癌临床分期有一定指导意义,并对预后判断具有重要价值.     

外周血循环肿瘤细胞在卵巢癌化疗中的临床应用

目的 探讨卵巢癌患者外周血中循环肿瘤细胞（CTC）变化情况与化疗疗效的关系。方法 选择2013年2月至2015年4月间在我院接受化疗的卵巢癌患者28例,分别抽取其化疗前后外周静脉血4 ml,采用免疫磁珠微粒阴性富集技术和免疫荧光原位杂交技术（imFISH）检测CTC数目,以CTC≥2个/3.2 ml定义为阳性。采用RECIST 1.0版标准评价化疗疗效,观察化疗前后CTC变化与疗效的关系。结果 化疗前外周血CTC阳性率为60.71％（17／28）,其与肿瘤分期（P=0.019）及CA125水平（P=0.022）有关;化疗后CTC阳性率为25％（7／28）。根据化疗前后CTC的变化,将患者分为阳性 阳性、阳性 阴性、 阴性 阳性、阴性 阴性4组,4组有效率分别为80.0％（4／5）、83.3％（10／12）、0（0／2）和88. 9％（8／9）,差异有统计学意义（P=0.045）。结论 化疗前后CTC变化与疗效有一定关系,CTC可作为评估卵巢癌化疗疗效的参考指标。     

肺癌循环肿瘤细胞的临床意义

转移是导致肺癌患者死亡的首因,而循环肿瘤细胞（Circulating tumor cells,CTCs）在形成远处转移过程中起关键作用。近年研究显示,CTCs可能成为肺癌诊疗的新兴标志物及治疗新靶点,本文从诊断与临床分期、转移与复发、个体化治疗与预后等方面阐述肺癌CTCs的临床意义,以期为肺癌治疗提供新策略。     

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.     

Circulating Tumor Cells in Metastatic Breast Cancer：Monitoring Response to Chemotherapy and Predicting Progression-Free Survival

Objective  

The purpose of this study is to explore RT-PCR method to set up the examination platform for detecting circulating tumor cells (CTC) in peripheral blood from metastatic breast cancer patients. The primary endpoint is to find out the correlation of existence of CTC with clinical responses and progression-free survival (PFS).     

Circulating tumor cells (CTCs): Detection methods and their clinical relevance in breast cancer

The enumeration of circulating tumor cells has long been regarded as an attractive diagnostic tool, as circulating tumor cells are thought to reflect aggressiveness of the tumor and may assist in therapeutic decisions in patients with solid malignancies.However, implementation of this assay into clinical routine has been cumbersome, as a validated test was not available until recently. Circulating tumor cells are rare events which can be detected specifically only by using a combination of surface and intracellular markers, and only recently a number of technical advances have made their reliable detection possible. Most of these new techniques rely on a combination of an enrichment and a detection step.This review addresses the assays that have been described so far in the literature, including the enrichment and detection steps and the markers used in these assays. We have focused on breast cancer as most clinical studies on CTC detection so far have been done in these patients.     

循环肿瘤细胞检测在胃癌中的临床应用

循环肿瘤细胞（CTC）为存在于肿瘤患者外周血的一类特殊细胞,目前 CTC 检测已应用于胃癌患者生存期预测、术后复发的检测及个体化治疗指导等方面。随着研究的不断深入,CTC 将为胃癌的综合治疗提供新的帮助。     

Progress of circulating tumor cells in lung cancer

Circulating tumor cells (CTCs) are present in the peripheral blood, and play an important role in the recurrence and metastasis of lung cancer. As the researches about CTCs and targeted therapy move along, it is found that CTCs can be used as a "liquid" tumor tissue to guide clinical treatment and have a certain significance in the clinical staging and prognosis evaluation of lung cance.