循环肿瘤细胞检测在肺癌中的应用

肺癌是目前全世界范围内发病率和病死率最高的恶性肿瘤,早期诊治是改善其预后的关键,随着循环肿瘤细胞(CTCs)检测技术的改进与发展,检测的灵敏度与特异度逐步提高,肺癌患者将在早期得到确诊,对肺癌的诊断、治疗及预后评估都具有重要意义。本文重点综述了近年来CTCs检测诊断技术的进展及其在肺癌诊断中的应用,并对其检测技术面临的挑战进行了探讨。     

循环肿瘤细胞检测在肺癌中的研究进展

肺癌是我国最常见的恶性肿瘤之一,血清肿瘤标志物、影像学以及病理学检查是目前肺癌的主要检查手段,但这些检查方式在肺癌的临床应用中都具有自身的局限性。循环肿瘤细胞(circulating tumor cells,CTCs)属于"液体活检"中的一种,在肺癌的临床分期、疗效评价、早期诊断、复发及转移的监测方面都有着重要的作用。此外,对于不能获得肿瘤组织的晚期肺癌患者,CTCs因其来源于肿瘤原发灶而可作为一种替代途径指导个体化治疗。该文就CTCs在肺癌中的研究进展作一综述。     

循环肿瘤细胞分型检测及其在晚期肺癌中的临床应用研究

目的 对晚期肺癌患者外周血循环肿瘤细胞(CTCs)的数量和分型进行检测、分析,探讨其与患者临床特征的关系.方法 收集43例晚期肺癌患者的临床资料.基于CanPatrol?CTCs分型检测平台,通过RNA原位杂交技术及分子DNA技术荧光标记特异信使RNA(mRNA),将患者外周血中CTCs分为上皮型、混合型、间质型3种亚...     

循环肿瘤细胞检测及其特性

肿瘤转移是导致肿瘤患者死亡的重要原因,现有影像学手段及传统肿瘤标志物不能辅助医生尽早发现肿瘤转移并判断预后。外周血循环肿瘤细胞（circulating tumor cells,CTCs）存在于肺癌、乳腺癌、前列腺癌、结直肠癌、膀胱癌和卵巢癌等多种恶性肿瘤中,     

循环肿瘤细胞与肿瘤标志物在肺癌诊断中的价值比较

目的探讨循环肿瘤细胞(CTCs)检测在肺癌中的诊断价值。方法选取35例肺癌患者为肺癌组,以同期入院的非结核肺良性疾病患者22例及健康体检者26例为对照组,分别检测两组患者外周血CTCs及传统血清肿瘤标志物的表达水平,并使用灵敏度、特异度及ROC曲线对CTCs在肺癌中的诊断价值进行评价。结果肺癌组CTCs的检出率为77.1%,显著高于对照组CTCs检出率(10.4%),差异具有统计学意义(P<0.05);CTCs诊断肺癌的灵敏度、特异度、ROC曲线下面积分别为77.1%、89.6%、0.865,均高于传统血清肿瘤标志物NSE(42.9%、77.1%、0.591),CA125(34.3%、87.5%、0.706),CA153(11.4%、89.5%、0.530),CYFRA21-1(62.9%、64.6%、0.674),SCCA(22.9%、66.7%、0.336);而CEA的特异度(93.8%)虽高于CTCs,但其灵敏度(45.7%)及ROC曲线下面积(0.793)均低于CTCs。结论CTCs检测可用于肺癌的诊断,且与传统血清肿瘤标志物相比具有更高的诊断价值。     

循环肿瘤细胞的检测及其在乳腺癌中的应用研究进展

现已证实,在乳腺癌患者确诊之前,就可能已经发生了血液转移,循环肿瘤细胞(CTCs)从原发肿瘤组织中脱落并进入血液循环,随血液循环到达远处组织并种植、生长,便形成转移病灶。如果能在早期检测出CTCs并描述其表型和基因型特征将促进乳腺癌个体化治疗方案的制订及治疗效果的判定,目前敏感的检测技术使得在单细胞水平检测和评价CTCs成为可能,该篇综述将讨论CTCs检测技术及在浸润性乳腺癌中的应用价值。     

液体活检技术在肺癌治疗中的应用

肺癌为全球肿瘤相关死亡的首要原因,分子靶向治疗药物为其治疗提供了新的选择,然而分子靶向治疗应以必要的基因检测为指导。传统的穿刺或组织活检技术依赖的基因检测,风险大,不宜反复进行。而液体活检以外周血、唾液等为样本,进行循环肿瘤细胞(CTC)、循环肿瘤核苷酸(ct DNA)检测,具有创伤小、可重复性、均化异质性、实时判断疗效等优势,在肺癌的诊断与分期、疗效与预后预测、转移与复发风险评估、肿瘤异质性与耐药性研究等方面具有重要应用前景。本文就近年来液体活检技术在肺癌领域的研究进展进行综述,并就CTC与ct DNA技术的优缺点、互补性以及目前的局限性进行简要讨论。     

循环肿瘤细胞的检测和应用

循环肿瘤细胞计数可以反映原发肿瘤的侵袭程度,已经成为实体肿瘤远处转移的一个早期诊断指标,与某些肿瘤的预后也存在一定相关性。最近发展的循环肿瘤细胞检测技术采用优化的细胞表面或/和细胞内标志物组合,可以更准确地筛选并富集目的细胞,提高了检测准确性和效率。     

非小细胞肺癌循环肿瘤细胞检测技术与临床应用的研究进展

转移和复发被认为是影响恶性肿瘤预后的主要因素,对于转移与复发形成的相关基础理论中,种子和土壤假说认为,原发肿瘤在生长过程中,有微小肿瘤细胞个体或集团通过循环系统的运送到达远处器官,如同种子播散在土壤中,形成与原发灶特性极其类似的转移灶,这一理论非常形象地说明了恶性肿瘤的转移途径,并点明种子肿瘤细胞即循环肿瘤细胞(CTCs)是转移和复发的关键,并有相关可靠的临床研究依据。近年来CTCs又引起肿瘤研究者的热情,将CTCs作为潜在肿瘤生物学标志物及研究肿瘤转移过程成为研究的热点。在本综述中,我们将对CTC生物学和转移相关基因检测、现今最常用的检测技术,及其在临床研究尤其在肺癌领域中的应用进行阐述。     

循环肿瘤细胞研究新进展

恶性肿瘤释放癌细胞进入血液、形成远处转移灶,导致治疗效果差、预后不良。如果在转移灶形成之前能够在血液中发现循环肿瘤细胞(CTCs),则可早期发现转移灶,在治疗期间通过外周CTCs检测来观察治疗效果和判断预后。通过检测外周CTCs的凋亡和上皮间质转变,以及对循环肿瘤干细胞(CSCs)特性的研究,可以为临床治疗提供有效数据,因此,能否有效地检测出外周血CTCs以及研究其特性直接关系到恶性肿瘤的治疗和预后。本文就外周CTCs在恶性肿瘤中的研究进展做一综述。     

循环肿瘤细胞的磁分离法建立及在肺癌EGFR-TKI治疗中的应用

目的探索磁分离循环肿瘤细胞(circulating tumor cells,CTCs)联合竞争性等位基因特异性Taq ManPCR(Cast PCR)检测表皮生长因子受体(EGFR)基因突变的可行性。方法收集12例晚期非小细胞肺癌(non-small cell lung cancer,NSCLC)患者血液样本各7.5 m L,两步法磁分离CTCs,免疫荧光染色检测上皮细胞黏附分子(Ep CAM)和细胞角蛋白(Cytokeratin)后,流式细胞术检测CTCs数量及纯度;Cast PCR检测CTCs及循环DNA中EGFR基因del EGFR19、T790M和L858R突变;用EGFR基因突变检测试剂盒(ADx-ARMS法)检测癌组织的上述位点突变。结果高表达与不表达Ep CAM的CTCs分别在6例和12例NSCLC患者中被检测出;del EGFR19、T790M和L858R突变例数分别在2例、8例和5例NSCLC患者的CTCs中被检测出,其总检出率为83.3%(10/12);2例患者的循环DNA中检出L858R突变;ADx-ARMS法检测12例NSCLC患者癌组织中上述3种突变分别是2例、5例和3例,总检出率为75.0%(9/12)。CTCs与癌组织的突变具有一致性(Kappa=0.75,P=0.007),循环DNA与癌组织的突变一致性无统计意义(Kappa=0.06,P=0.546)。结论磁分离CTCs联合Cast PCR能有效检测EGFR基因突变,值得在NSCLC患者的EGFR酪氨酸激酶抑制剂(EGFR-TKI)治疗中推广应用。     

Role of circulating tumor cells in diagnosis of lung cancer: a systematic review and meta-analysis

ObjectiveWe systematically reviewed the literature relating to the diagnostic accuracy of circulating tumor cells (CTCs) for the clinical determination of lung cancer.MethodsThis meta-analysis aimed to evaluate the diagnostic accuracy of CTCs for the clinical determination of lung cancer. The PubMed, Embase, Cochrane Library, and Web of Science databases were searched for relevant studies up to 31 May 2020. The numbers of patients with true positive, false positive, false negative, and true negative results were extracted from each individual study. Pooled sensitivity, specificity, and area under the curve values were calculated with 95% confidence intervals (CI).ResultsTwenty-one studies with 3997 subjects met the inclusion criteria. The overall diagnostic accuracy was assessed. The pooled sensitivity and specificity were 0.72 (95%CI: 0.65–0.79) and 0.96 (95%CI: 0.91–0.98), respectively, and the pooled positive and negative likelihood ratios were 16.86 (95%CI: 7.65–37.12) and 0.29 (95%CI: 0.23–0.37), respectively. The combined diagnostic odds ratio was 58.12 (95%CI: 24.82–136.09).ConclusionThis meta-analysis indicated that CTCs had good diagnostic value for detecting lung cancer.     

Standardized quantification of circulating peripheral tumor cells from lung and breast cancer.

Detection and quantitation of circulating tumor cells from solid epithelial tumors could become a valuable tool for therapy monitoring if the procedure can be standardized. In the present work we assessed the influence of pre-analytical handling, storage and white blood cell isolation on analysis of a population of spiked tumor cell-line cells and intrinsically present epithelial cells in the peripheral blood of breast and lung cancer patients and the sensitivity of their detection. Sucrose density separation did not enrich epithelial cells, and even depleted them, leading to a gross underestimation of their numbers (3/13 positive, between 2.9 and 50 cells/mL) in comparison to red blood cell lysis (13/13 positive, between 77,200 and 800 cells/mL). Short-term storage of whole blood samples for up to 7 days had little influence on the number of epithelial cells recovered. The effectiveness of magnetic bead enrichment was dependent on the number of relevant cells and the volume used for enrichment. Red blood cell lysis and fluorochrome-labeled antibody staining in a no-wash procedure with subsequent laser scanning cytometry allowed the detection of circulating epithelial cells in 92% of breast and lung cancer patients. Two examples of how this method can be applied for the longitudinal analysis in individual patients are shown, with an increase in numbers preceding relapse and a decrease paralleling tumor reduction. The proposed simple and easy method allows close monitoring, which may help in real-time analysis of the response of solid tumors, especially their systemic component, to therapy and hopefully will contribute to more individually tailored therapy.     

循环肿瘤细胞在中晚期非小细胞肺癌患者联合治疗中的疗效评价

目的分析循环肿瘤细胞(CTCs)在中晚期非小细胞肺癌(NSCLC)患者化疗联合分子靶向治疗过程中的疗效评价能力。方法采用回顾性研究方法,选取2014年8月至2018年2月河北省涿州市医院收治的采用吉西他滨加顺铂(GP)化疗、联合吉非替尼靶向治疗的中晚期NSCLC患者共33例,共观察4个疗程。分别于治疗前,以及治疗2、4个疗程后按照实体肿瘤疗效评价标准(RECIST)检测肿瘤直径及肿瘤标志物变化;并通过免疫磁珠法富集、Envision法检测外周血CTCs的变化,分析CTCs标准与RECIST标准的异同。结果联合治疗前,33例中晚期NSCLC患者均检测出CTCs,检测阳性率为100%,而基线CTCs数量在不同年龄、性别、肿瘤类型和临床分期和吸烟史患者间差异无统计学意义(P>0.05);而肿瘤直径越大,CTCs数量越多(P<0.05)。Spearman相关性分析显示,第2疗程后CTCs变化与治疗效果具有相关性(r=0.832,P=0.000)。按照RECIST标准分为有效组(n=19)和无效组(n=14),有效组治疗第2疗程和第4疗程后CTCs计数分别为8.0(3.0,15.0)个/5 ml及5.0(3.0,10.0)个/5 ml与治疗前12.0(7.0,18.0)个/5ml相比,差异均具有统计学意义(Z=-3.427、-3.876,P<0.05);无效组治疗第2疗程和第4疗程后CTCs计数分别为16.5(12.0,20.0)个/5 ml,18.5(14.0,22.0)个/5 ml与联合治疗前14.5(10.0,18.0)个/5 ml相比,差异均具有统计学(Z=-3.126、-2.897,P<0.05)。采用Fisher’s确切概率法评价两种疗效判定方法,两者差异无统计学意义(P>0.05)。结论中晚期NSCLC联合治疗过程中,CTCs测定可以实现早期、有效的疗效评价,是RECIST标准的有益补充。     

A combination of circulating tumor cells and CA199 improves the diagnosis of pancreatic cancer

BackgroundEarly diagnosis of pancreatic ductal adenocarcinoma (PDAC) is difficult due to the lack of effective screening tests. CA199, the standard biomarker for PDAC management, is not sufficiently reliable for early diagnosis. This prospective study aimed to evaluate whether circulating tumor cells (CTCs) could complement or perform better than CA199 in determining PDAC.MethodsA total of 168 blood samples were collected from 80 patients with PDAC, 32 patients with acute pancreatitis, 22 patients with benign pancreatic masses, and 34 healthy donors. CTCs were detected by a novel system combining negative enrichment with immunostaining and fluorescence in situ hybridization (NE‐imFISH). Next, ROC curves and AUC analyses were conducted to assess diagnostic abilities of CA199, CTCs, and the combination of the two biomarkers in PDAC.ResultsCTCs were stained as CD45–/DAPI+/CEP8 ≥3. With 2 CTCs/3.2 ml as the cut‐off value, the sensitivity/specificity of the CTC number was 0.76/0.94, which was comparable to that of CA199 (0.78/0.83; Delong test p = 0.3360). Improved performance was achieved through a logistic regression model integrating CA199 and CTC number (AUC_CTC+CA199 = 0.95, AUC_CA199 = 0.80, AUC_{CTC number} = 0.85; Delong test p _vs. _CA199 < 0.0001 and p _vs. _{CTC number} = 0.0002). CTC subtype was inferior to CTC number as a diagnostic marker (AUC_{CTC subtype} = 0.73; Delong test p _vs. _{CTC number} < 0.0001).ConclusionThe dual‐marker panel consisting of CA199 and CTC number can significantly improve upon the diagnostic performance of CA199 alone, highlighting the promising clinical utilization as an effective strategy for PDAC surveillance.     

外周血液中循环肿瘤细胞和循环游离DNA检测在乳腺癌患者中的应用

目的 探究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应用效果.方法 收集本院2018-02-2019-02收治的94例乳腺癌患者、48例乳腺良性疾病患者及56例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...