循环肿瘤细胞核型对非肌层浸润性膀胱癌预后的预测价值

  目的  探究外周血中循环肿瘤细胞 (circulating tumor cell，CTC) 和循环肿瘤细胞核型对非肌层浸润性膀胱癌（non-muscle invasive bladder cancer，NMIBC）患者预后的预测作用。  方法  选取2018年5月至2019年5月于承德医学院附属医院首次诊治的109例患者临床资料，其中NMIBC组58例、良性病变组51例。采用差相富集和免疫荧光染色-染色体荧光原位杂交（SE-iFISH)技术行CTC鉴定及核型分析，以患者CTC三倍体数目≥60%为三倍体组，数目<60%为非三倍体组，并行生存和预后分析。  结果  NMIBC组患者的CTC阳性率为81.0%（47/58），显著高于良性病变组的9.8%（5/51）。Kaplan-Meier法和Log-rank法分析显示，术前、术后三倍体组患者的无病生存（disease-free survival，DFS）期明显短于非三倍体组(P<0.001)，术后CTC阳性患者的DFS短于CTC阴性患者（P＜0.05）。行Cox比例风险回归模型多因素分析显示术前、术后CTC核型是影响NMIBC患者预后的独立危险因素。  结论  术前、术后监测CTC和CTC核型对NMIBC患者具有预测作用，CTC核型是影响预后的独立因素。      

循环肿瘤细胞与乳腺癌患者预后相关性的Meta分析

  目的  评估循环肿瘤细胞（circulating tumor cell,CTC）在预测乳腺癌患者预后中的作用。  方法  检索Medline、Embase、中国数字化期刊全文数据库（CNKI）、万方数据库及维普全文网中国内外相关文献,收集检测外周血肿瘤细胞在乳腺癌患者预后中的研究,使用Review Manager 5.1.4中的方差倒数法对无进展生存期（progression-free survival,PFS）和总生存期（overall survival,OS）进行Meta分析。  结果  共纳入33篇英文文献,总计5 393例乳腺癌患者。Meta分析结果提示CTC阳性组较CTC阴性组在PFS [HR=2.09（95%CI:1.72~2.55）,n=23,I2=57%]和OS[HR=2.49（95%CI:2.18~2.85）,n=24,I2=0]方面差异均有统计学意义（P < 0.01）。根据UICC（International Union Against Cancer）肿瘤分期进行亚组分析,结果显示CTC对不同期别肿瘤的PFS和OS也均有预测价值（P < 0.01）。  结论  外周血CTC阳性的乳腺癌患者与CTC阴性患者相比预后较差,CTC可以作为乳腺癌患者的预后预测指标。      

尿路上皮癌患者外周血循环肿瘤细胞检测及其与临床特征和预后的关系

目的:探究尿路上皮癌（urothelial carcinoma,UC）患者手术前、手术后外周血循环肿瘤细胞（circulating tumor cell,CTC）与临床特征和预后的关联。方法:回顾性收集2018年09月至2019年11月我院诊治的87例患者,其中57例为UC组,30例为尿路上皮的良性疾病组,于术前和术后分别两次对UC组进行外周血CTC检测,良性疾病组仅术前进行CTC检测,并分析CTC与患者的临床特征及无进展生存期（progression-free survival,PFS）的关联。结果:UC组中CTC检测阳性率为84.2%（48/57）,与良性疾病组比较差异具有统计学意义（P＜0.001）。CTC与性别、T分期、血尿、肿瘤直径及肿瘤浸润深度相关,而与年龄、分化程度、病理类型以及病灶数无关。Log-rank检测显示,术后CTC与患者的PFS密切相关（P=0.001）,而术前CTC与预后无关。Cox比例风险回归模型提示,T分期及术后CTC是影响患者PFS的独立危险因素。结论:术后外周血CTC是UC患者不良预后的独立危险因素,CTC可能成为诊断疾病和监测患者预后有力的生物标志物。     

术前红细胞分布宽度对非小细胞肺癌患者预后的评估价值

  目的  分析术前红细胞分布宽度（red cell distribution width,RDW）对非小细胞肺癌（non-small cell lung cancer,NSCLC）患者预后的评估价值。  方法  回顾性分析2008年3月至2012年12月天津医科大学肿瘤医院行根治性手术的513例NSCLC患者的临床病理资料。根据受试者工作特征ROC曲线分析确定RDW的截断值。分别通过χ2、t或秩和检验分析RDW与临床病理特征和实验室变量之间的关系,采用Kaplan-Meier法进行生存分析,Log-rank检验组间生存差异。采用Cox比例风险回归模型进行多因素分析。  结果  根据ROC曲线,术前RDW最佳截断值为12.95%。其中RDW≤12.95%为250例,RDW>12.95%为263例。术前RDW与年龄、血红蛋白、白蛋白、纤维蛋白原和D-二聚体水平显著相关（均P < 0.05）。单因素分析显示肿瘤位置、病理分期、血小板数目、白蛋白、血红蛋白、纤维蛋白原和术前RDW是影响NSCLC患者预后的因素（均P < 0.05）;多因素分析显示术前RDW和病理分期是影响NSCLC患者预后的独立危险因素（均P < 0.05）。  结论  术前RDW水平可作为预测行根治性手术的NSCLC患者预后的指标。      

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。     

胃癌患者循环肿瘤细胞的检测及临床意义

  目的  探讨胃癌患者外周血中循环肿瘤细胞的临床意义。  方法  采集45例胃癌患者的外周血,应用密度梯度离心法和免疫荧光法检测外周血中的循环肿瘤细胞(circulating tumor cells,CTCs),并比较其临床病理意义。  结果  应用密度梯度离心法和免疫荧光法检测胃癌患者外周血中的CK19阳性肿瘤细胞即为CTCs。45例胃癌患者中27例为CTCs阳性,CTCs阳性与淋巴结转移(P=0.007)、远处转移(P=0.035)和复发密切相关(P=0.035),而与性别、年龄、肿瘤部位和TNM分期、肿瘤分化程度无关(P> 0.05)。  结论  CTCs与胃癌的不良预后相关。      

基于核酸适配子的循环肿瘤细胞检测研究进展

循环肿瘤细胞（circulating tumor cell,CTC）是从肿瘤病灶脱落后进入血液循环的肿瘤细胞,在肿瘤的微转移检测、病情评估、疗效及预后评判等方面具有重要价值,但因其在血液中数量极少,难以检测,临床应用进展缓慢。近年来,随着生物学技术的不断发展,CTC的富集及检测新方法不断进步,其中基于核酸适配子的富集及检测方法不但能快速高效捕获和无损释放CTC,还能定性、定量分析CTC,甚至能检测出血液中的单个CTC和不同亚群的肿瘤细胞,具有良好的应用前景。      

食管癌放化疗中循环肿瘤细胞的变化及其临床意义

目的:探讨局部晚期食管癌放化疗中循环肿瘤细胞（circulating tumor cells ,CTCs）的动态变化,以及CTCs与食管癌临床特征和预后的相关性分析。方法:利用免疫磁珠富集联合免疫荧光检测2011年5 月至2013年5 月江苏大学附属人民医院48例局部晚期食管癌患者放化疗前后的循环血肿瘤细胞,对比分析放化疗前后的动态变化,并分析循环肿瘤细胞与患者临床特征及2 年生存率的关系。结果:循环血肿瘤细胞与肿瘤侵犯程度、淋巴结转移状态及临床分期均显著相关（P < 0.05）,治疗前循环血肿瘤细胞阳性率为52.1%（25/ 48）,治疗结束后为20.8%（10/ 48）,差异具有统计学意义（P < 0.01）。 放化疗前后的循环肿瘤细胞均与患者的2年生存率密切相关（P < 0.05）,Cox 回归分析提示临床分期及放化疗后循环血肿瘤细胞水平是食管癌患者的独立预后因子。结论:循环肿瘤细胞可以反映局部晚期食管癌患者的疾病进展程度,并且可以作为判断预后的指标。      

DC-CIK对结直肠癌根治术后肝转移患者疗效和循环肿瘤细胞的影响

目的: 探讨树突状细胞-细胞因子诱导的杀伤细胞（dendritic cell-cytokine induced killer cell, DC-CIK）对结直肠癌（colorectal carcinoma, CRC）根治术后肝转移患者循环肿瘤细胞（circulating tumor cell, CTC）数量、疗效和预后的影响。方法: 回顾性分析2009 年7 月至2015 年12 月在解放军第309 医院普通外科采用DC-CIK+常规疗法治疗的CRC根治术后肝转移患者62例（DC-CIK组）和同期未接受DC-CIK治疗的70 例患者（常规组）的临床资料,同时抽取部分患者（DC-CIK组21 例,常规组24 例）的外周静脉血,用CellSearch R 免疫磁珠技术检测血中CTC的数量,分析比较两组患者的疗效和预后。结果：DC-CIK 组治疗后患者CTC数量明显低于治疗前[ （1.0±1.1）vs（2.7±2.0）个,P<0.01],而常规组CTC变化不明显（P>0.05）;DC-CIK组肝转移灶手术切除率、治疗客观有效率、患者无进展生存和总生存均显著优于常规组（P<0.05 或P<0.01）。结论：DC-CIK可减少CRC根治术后肝转移患者CTC数量并延长患者生存时间,在规范化实施的前提下具有可靠的抗肿瘤效果。     

微流控芯技术检测循环肿瘤细胞在脑胶质瘤预后预测中的价值

背景与目的：脑胶质瘤患者的预后既往多通过生化指标进行预测,临床实践中有一定难度且受多种因素影响,导致预测效率相对较差。探讨微流控芯技术检测循环肿瘤细胞（circulating tumor cell,CTC）在脑胶质瘤预后预测中的价值。方法：选取2016年3月—2020年3月内蒙古自治区人民医院神经外科收治的96例脑胶质瘤患者为研究对象,采用微流控芯技术富集患者外周血CTC,并利用免疫荧光法对CTC进行鉴定;采用logistic回归方程分析脑胶质瘤患者外周血CTC的检测结果与脑胶质瘤临床特征之间的关系并建立受试者工作特征（receiver operating characteristic,ROC）曲线;制作并分析脑胶质瘤患者术后生存曲线。结果：微流控芯技术对CTC富集联合免疫荧光结果显示,CTC检测阳性率为42.71%,外周血检测出的CTC数目为（66.27±6.36）/mL;单因素分析结果显示,外周血CTC与患者的肿瘤分期、组织学类型、淋巴结转移是否存在囊性病变、神经元特异性烯醇化酶（neuron-specific enolase,NSE）密切相关（P<0.05）,与年龄、性别、KPS评分、血清胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）含量等临床特征无相关性;多因素分析结果显示肿瘤分期、组织学类型及NSE的表达与CTC密切相关;通过ROC曲线分析得出肿瘤分期、组织学类型以及NSE的AUC分别为0.645、0.687、0.720,预测概率AUC为0.814（P<0.05）;CTC阳性组和阴性组患者1年生存率差异无统计学意义（P>0.05）,但两组患者2年和3年生存率差异有统计学意义（P<0.05）。结论：微流控芯技术可定量检测脑胶质瘤患者外周血CTC,其水平与肿瘤分期、组织学类型、NSE密切相关,直接影响脑胶质瘤患者远期生存率。因此,外周血CTC可作为脑胶质瘤患者预后预测的重要指标。     

Combined detection and subclass characteristics analysis of CTCs and CTECs by SE-iFISH in ovarian cancer

ObjectiveHematogenous metastasis is essential for the progression of ovarian cancer (OC), and circulating tumor cells (CTCs) are part of the metastatic cascade. However, the detection rate of CTC is low due to the use of less sensitive detection methods. Therefore, this study aimed to detect CTCs and circulating tumorigenic endothelial cells (CTECs) in patients with OC using subtraction enrichment and immunostaining and fluorescence in situ hybridization (SE-iFISH). MethodsWe enrolled a total of 56 subjects, including 20 OC patients and 36 ovarian benign tumor patients. CTCs and CTECs were captured by subtraction enrichment (SE) and counted and classified according to immunofluorescence staining of tumor markers (TMs) carbohydrate antigen 125 (CA125) and human epididymis protein 4 (HE4) combined with fluorescence in situ hybridization (iFISH) of chromosome 8 (Chr8) aneuploidy. The diagnostic value and subtype characteristics of CTCs and CTECs were investigated. ResultsThe detection rate of CTCs by SE-iFISH was high. Compared with CA125 and HE4, Chr8 aneuploidy was the major identification feature of CTC. CTC counts in OC were statistically higher than those in benign groups. CTC and CTEC with ≥pentaploidy were detected in both groups, illustrating the poor diagnostic value of CTC or CTEC. Distributions of triploid and tetraploid CTC subtypes were significantly different, and combined detection of triploid and tetraploid CTCs showed the best diagnostic value. In contrast, the distribution of CTECs in the OC and benign groups had no statistically significant difference. Small CTCs accounted for over 1/3 of the total CTC count. We also found that small CTCs and CTECs primarily comprised triploid cells, while large CTCs and CTECs mainly comprised pentaploidy and beyond.ConclusionsThe application of SE-iFISH offered a more comprehensive understanding of heterogeneous CTCs and CTECs in OC. Analysis of subclass characteristics of the CTCs and CTECs according to Chr8 aneuploidy and cell size may broaden their potential clinical utility and deepen mechanistic studies in OC.     

Prognostic significance of circulating tumour cells enumerated after filtration enrichment in early and metastatic breast cancer patients

SummaryBackground We previously found a higher incidence of circulating tumour cells (CTCs) in women with metastatic breast cancer compared to early disease. In this study, we present follow-up data to explore the prognostic significance of these findings.Methods CTCs were quantified by immunostaining and direct visualization after centrifugation and filtration enrichment of peripheral blood from 131 patients. Time to progression (TTP) and overall survival (OS) were defined as interval from first blood sampling to first documented disease progression, or death respectively. Lifetime data was analysed using Kaplan–Meier method, log-rank test and Cox proportional hazards model.Results Follow-up data is available for 123 patients. In early disease, median CTC≥4 best distinguished patients with shorter TTP (p=0.05, log-rank test). In univariate analysis, tumour size, grade, lymphovascular invasion (LVI) and receptor status significantly related to TTP but none of the covariates related to OS. In multivariate analysis, T stage was the only independent predictor of TTP. In metastatic disease, median CTC≥13 optimally identified patients with shorter TTP (p=0.01). In univariate analysis, median CTC level ≥13 and prior lines of chemotherapy predicted for TTP while in multivariate analysis, median CTC level ≥13 was the only significant independent prognostic factor (p=0.02). No relationship between CTC level and OS was found in this subgroup.Conclusion Median CTC level determined in the course of treatment predicts for TTP in metastatic breast cancer. In early breast cancer, an association was found between CTC level and TTP although this did not reach statistical significance (p=0.05).     

乳腺癌新辅助化疗中循环肿瘤细胞的动态变化及疗效观察

  目的  通过检测新辅助化疗乳腺癌患者化疗前及化疗后各周期外周血中循环肿瘤细胞的变化,比较TC及TEC两种不同新辅助化疗方案的疗效。  方法  选取2010年1月~2012年12月间本院收治的96例局部晚期乳腺癌患者,随机分为两组,分别给予TC（多西他赛+环磷酰胺）和TEC（多西他赛+表柔比星+环磷酰胺）两种方案4个周期。分别抽取新辅助化疗前、新辅助化疗1、2、3、4个疗程后48h患者外周血5 mL,并采用流式细胞术测量外周血中循环肿瘤细胞含量,比较两种新辅助化疗方案疗效。  结果  两组患者新辅助化疗前年龄、绝经状态、ER、PR、C-erbB-2、肿物大小、临床分期等一般资料无统计学差异;两组患者新辅助化疗前外周血中循环肿瘤细胞值无明显差异;新辅助化疗前,两组患者外周血中循环肿瘤细胞含量与肿物体积有关系,肿物>5 cm者循环肿瘤细胞含量明显高于肿物≤5 cm者（P均 < 0.05）;两种新辅助化疗方案化疗后,两组患者外周血中循环肿瘤细胞值随化疗周期进行而持续降低（P均 < 0.05）,同时,使用TEC方案患者外周血中循环肿瘤细胞值明显低于使用TC方案患者（P均 < 0.05）。  结论  使用TC和TEC方案新辅助化疗可使局部晚期乳腺癌患者病灶缩小,外周血中循环肿瘤细胞值降低。同时,监测外周血中循环肿瘤细胞值变化,可评价新辅助化疗方案疗效。      

Role of aneuploid circulating tumor cells and CD31+ circulating tumor endothelial cells in predicting and monitoring anti‐angiogenic therapy efficacy in advanced NSCLC

Prognosticating the efficacy of anti‐angiogenic therapy through longitudinal monitoring and early detection of treatment resistance in cancer patients remain highly challenging. In this study, co‐detection and comprehensive phenotypic and karyotypic molecular characterization of aneuploid circulating tumor cells (CTCs) and circulating tumor endothelial cells (CTECs) were conducted on non‐small cell lung cancer (NSCLC) patients receiving bevacizumab plus chemotherapy. Prognostic values of the cell‐based significant univariate risk factors identified by Cox regression analyses were progressively investigated. Subjects showing an increase in total post‐therapeutic platelet endothelial cell adhesion molecule‐1 (CD31)^– CTCs and CD31⁺ CTECs exhibited a significantly reduced median progression‐free survival (mPFS) and overall survival. Further stratification analyses indicated that pretherapeutic patients bearing vimentin (Vim)⁺ CTECs (mesenchymal M‐type) at baseline revealed a significantly shortened mPFS compared with patients with Vim^– CTECs. Post‐therapeutic patients harboring epithelial cell adhesion molecule (EpCAM)⁺ CTCs and CTECs (epithelial E‐type), regardless of Vim expression or not, showed a significantly reduced mPFS. Post‐therapeutic patients possessing de novo EpCAM⁺/Vim⁺ (hybrid E/M‐type) CTECs displayed the shortest mPFS. Patients harboring either pre‐ or post‐therapeutic EpCAM^–/Vim^– null CTECs (N‐type) exhibited a better response to therapy compared to patients harboring EpCAM⁺ and/or Vim⁺ CTECs. The presented results support the notion that baseline Vim⁺ CTECs and post‐therapeutic EpCAM⁺ CTCs and CTECs are predictive biomarkers for longitudinal monitoring of response to anti‐angiogenesis combination regimens in NSCLC patients.     

Clinical significance of circulating tumor cells in breast cancer patients

Circulating tumor cells (CTCs) are defined as tumor cells circulating in the peripheral blood of patients, shed from either the primary tumors or its metastases. Many techniques have been developed in the recent years to identify CTCs in breast cancer patients, and trials have proved the prognostic significance of CTCs. In this study, we validated the CTC detection method of combining cell filtration and laser scanning cytometry (LSC), which was highly reproducible with increased sensitivity and accuracy. In 134 non-metastatic breast cancer patients analyzed, HER2 was found to be the only primary tumor characteristics that correlated with the presence of CTCs. 85 patients with definitive stage information were selected for association study between the disease stages and CTC numbers. The detection rate and the number of CTCs were correlated with the disease stages. Moreover, assessment of CTCs in 92 metastatic breast cancer patients was found to be able to predict the efficacy of chemotherapy. Increase in CTC numbers was an independent prognostic factor for treatment outcomes. Our results suggested that CTC assessment could be an indication of the disease progression and analysis of the properties of CTCs is likely to provide new insights into the biology of breast cancer and contribute to defining novel treatments and better prediction of clinical outcomes.     

Detection rate and prognostic value of circulating tumor cells and circulating tumor DNA in metastatic uveal melanoma

Circulating tumor cells (CTCs) and circulating tumor DNA (ctDNA) have been recently investigated in several cancer types, but their respective clinical significance remains to be determined. In our prospective study, we compared the detection rate and the prognostic value of these two circulating biomarkers in patients with metastatic uveal melanoma. GNAQ/GNA11 mutations were characterized in archived tumor tissue. Using a highly sensitive and mutation‐specific bidirectional pyrophosphorolysis‐activated polymerization (bi‐PAP) technique, GNAQ c.626A>T, GNAQ c.626A>C and GNA11 c.626A>T copy numbers were quantified in plasma from 12 mL of blood. CTCs were detected at the same time in 7.5 mL of blood by the CellSearch® technique. Patient characteristics and outcome were prospectively collected. CTCs (≥1) were detected in 12 of the 40 included patients (30%, range 1–20). Among the 26 patients with known detectable mutations, ctDNA was detected and quantified in 22 (84%, range 4–11,421 copies/mL). CTC count and ctDNA levels were associated with the presence of miliary hepatic metastasis (p = 0.004 and 0.03, respectively), with metastasis volume (p = 0.005 and 0.004) and with each other (p < 0.0001). CTC count and ctDNA levels were both strongly associated with progression‐free survival (p = 0.003 and 0.001) and overall survival (p = 0.0009 and <0.0001). In multivariate analyses, ctDNA appeared to be a better prognostic marker than CTC. In conclusion, ctDNA and CTC are correlated and both have poor prognostic significance. CTC detection can be performed in every patient but, in patients with detectable mutations, ctDNA was more frequently detected than CTC and has possibly more prognostic value.