膜过滤分离肿瘤细胞技术研究
摘要
目的：利用体外细胞实验、动物实验及临床实验来对膜过滤分离肿瘤细胞技术(Isolation by size of epithelial tumor cells, ISET)进行系统性研究,目的是为了建立起一整套使用ISET技术在血液循环过程中检测、筛选、分离循环肿瘤细胞(circulating tumor cells, CTC)并进行深入分析的技术方法,并为进一步的临床试验打下基础。
研究方法：1.利用带EGFP报告基因的H520肺鳞癌细胞株,构建ISET技术体外细胞实验方法及技术流程,并观察ISET技术在生理盐水和外周血这两种不同介质中的肿瘤细胞回收率及敏感性、特异性。2.建立肿瘤细胞血行转移动物模型,利用ISET技术对动物外周血进行检测,寻找转移的肿瘤细胞。3.ISET外周血筛查装置的设计和建立,以及对96例肺部疾病患者外周血进行筛查,观察ISET技术应用于临床检测的敏感性和特异性,并对特殊病例进行外周血CTC的持续监视。4.ISET血液循环系统的设计、建立,应用于VX2兔肿瘤细胞血行转移模型,对比ISET外周血筛查装置和血液循环系统的敏感性,验证ISET血液循环系统的安全性。
结果：1.成功将pEGFP-N3质粒转染入H520肿瘤细胞中,体外实验发现ISET技术在生理盐水和外周血这两种不同介质中的肿瘤细胞回收率分别为97.57±1.33%、79.53±2.63%,敏感性测试中基本达到了1ml外周血中检测到1个肿瘤细胞的敏感性,由于阴性对照组中未发现目标细胞,表明ISET技术有很高的特异性。2.分别建立NOD-SCID小鼠肿瘤细胞血行转移模型和VX2兔肿瘤细胞血行转移模型,利用ISET技术在3只NOD-SCID荷瘤鼠体内检测出CTC,在17只VX2荷瘤兔中有3只实验兔检测出CTC。3.设计并建立ISET外周血筛查装置并应用于96例肺部疾病患者,肿瘤组CTC的检出率为2.99%,非肿瘤组CTC的检出率为0.00%。远处转移组CTC的检出率为50.00%,显著高于未远处转移组CTC的检出率0.00%(P<0.05)。4.对结肠癌肺、骨转移患者进行外周血持续监测,治疗后期发现其外周血中开始持续存在CTC及癌栓。5.设计并建立ISET血液循环系统并应用于VX2兔肿瘤细胞血行转移模型,发现利用ISET外周血筛查装置的CTC的检出率为17.65%,显著低于利用ISET血液循环系统的检出率82.35%(P<0.05)。
结论：1.ISET技术在体外实验中有较高的敏感性和特异性,且实验方法简单,不依赖任何大型设备,实验结果稳定,可重复性强。2.在国际上首次利用ISET技术在肿瘤细胞血行转移动物模型中检测到CTC,具有极高的特异性。3.ISET外周血筛查装置适用于临床筛查CTC且方便组装便于携带。4.ISET技术在临床实验中表现出极高的特异性,但是敏感性不佳。5.ISET技术在疗效监视、评估预后以及在肿瘤转移机制研究方面有巨大潜力。6.在国际上首次设计并建立ISET血液循环系统,可以显著提高ISET技术敏感性,并且直接减少循环血液中存在的CTC数量。
上述技术、方法的实施对肿瘤诊断、疗效监视提供了新的思路。
OBJECTIVE:Systematic study of isolation by size of epithelial tumor cells (ISET) methodology in vitro, animal experiments and clinical trials. To establish a kind of method of ISET in the process of blood circulation to detect, screen and separate circulating tumor cells. This pilot study will serve as a basis for a much larger clinical trial.
METHOD:1.The method for the technical processes of ISET technology in vitro established with H520-pEGFP cells. H520-pEGFP cells will mix into the physiological saline and the blood sample from healthy volunteers respectively; detect to ascertain the recovery, the sensitivity and the specificity by ISET.2.To establish an animal model of tumor cells metastasis in the blood circulating, and detect the CTCs in the peripheral blood of animals by ISET technology.3.To design an ISET screening device, detect the CTCs in the peripheral blood of96patients with pulmonary disease. To observe the sensitivity and specificity of ISET screening device used in clinical testing. To monitor the presence of CTCs in one patient's peripheral circulation undergoing standard chemotherapy and radiotherapy.4.To design an ISET hemoperfusion system, detect the CTCs in the blood circulating of rabbit with VX2tumor, observe and compare sensitivity and security of ISET screening device and ISET hemoperfusion system.
RESULTS:1.The pEGFP-N3plasmid was transfected into H520tumor cells successfully. The recovery rate of mixed tumor cells detected by ISET technology remained about97.57±1.33in physiological saline and79.53±2.63in blood sample from healthy volunteers. We could detect1tumor cell in1ml peripheral blood while no positive cells were detected in healthy control, showing the high sensitivity and specificity of the ISET technology.2.The NOD-SCID mice model and rabbit model of tumor cells metastasis in the blood circulating have been established respectively.3samples in3NOD-SCID mice and3samples in17rabbit were detected with the CTCs by ISET technology.3.The ISET screening device was designed and built successfully to detect the CTCs in the peripheral blood of96patients with pulmonary disease.2samples in67cancer patients and0sample in29non-neoplastic patients were detected with the CTCs.2samples in4patients of stage IV and0sample in63patients of stage Ⅰ-Ⅲ were detected with the CTCs. However, CTCs were not detected in all6healthy samples by control. The detection rate of CTCs in4patients of stage IV was higher than that in63tumor patients of stage Ⅰ-Ⅲ (P<0.05).4.The persistence of CTCs in peripheral circulation in the patient of stage IV were monitored by ISET screening device.5.The ISET hemoperfusion system was designed and built successfully to detect the CTCs in the blood circulating of rabbit with VX2tumor. The detection rate of CTCs by ISET hemoperfusion system was higher than that by ISET screening device (P<0.05),82.35and17.65respectively (p<0.05).
Conclusion:1.The ISET technology to detect CTCs was convenient and stable with high sensitivity and good specificity. It does not rely on any large-scale experimental equipment.2.CTCs have been detected in animal models of tumor cells metastasis in the blood circulating at the first time in the international, indicating that ISET technology has a high specificity.3.ISET screening device suitable for clinical detecting CTCs and easy to assemble and carry.4.The ISET technology exhibit high specificity in clinical trials, but the sensitivity is poor.5.We can evaluate efficacy of chemotherapy and radiotherapy and other cancer-targeting therapies by using ISET technology that has great potential for mechanistic studies of ISET-isolated CTC biology.6.A new ISET hemoperfusion system could be designed to increase sensitivity to CTCs greatly and reduce the number of CTCs directly during the blood flow at the first time in the international.
The ISET hemoperfusion system gives us new insights into the diagnosis of malignant tumor and monitoring adjuvant therapy.
研究方法：1.利用带EGFP报告基因的H520肺鳞癌细胞株,构建ISET技术体外细胞实验方法及技术流程,并观察ISET技术在生理盐水和外周血这两种不同介质中的肿瘤细胞回收率及敏感性、特异性。2.建立肿瘤细胞血行转移动物模型,利用ISET技术对动物外周血进行检测,寻找转移的肿瘤细胞。3.ISET外周血筛查装置的设计和建立,以及对96例肺部疾病患者外周血进行筛查,观察ISET技术应用于临床检测的敏感性和特异性,并对特殊病例进行外周血CTC的持续监视。4.ISET血液循环系统的设计、建立,应用于VX2兔肿瘤细胞血行转移模型,对比ISET外周血筛查装置和血液循环系统的敏感性,验证ISET血液循环系统的安全性。
结果：1.成功将pEGFP-N3质粒转染入H520肿瘤细胞中,体外实验发现ISET技术在生理盐水和外周血这两种不同介质中的肿瘤细胞回收率分别为97.57±1.33%、79.53±2.63%,敏感性测试中基本达到了1ml外周血中检测到1个肿瘤细胞的敏感性,由于阴性对照组中未发现目标细胞,表明ISET技术有很高的特异性。2.分别建立NOD-SCID小鼠肿瘤细胞血行转移模型和VX2兔肿瘤细胞血行转移模型,利用ISET技术在3只NOD-SCID荷瘤鼠体内检测出CTC,在17只VX2荷瘤兔中有3只实验兔检测出CTC。3.设计并建立ISET外周血筛查装置并应用于96例肺部疾病患者,肿瘤组CTC的检出率为2.99%,非肿瘤组CTC的检出率为0.00%。远处转移组CTC的检出率为50.00%,显著高于未远处转移组CTC的检出率0.00%(P<0.05)。4.对结肠癌肺、骨转移患者进行外周血持续监测,治疗后期发现其外周血中开始持续存在CTC及癌栓。5.设计并建立ISET血液循环系统并应用于VX2兔肿瘤细胞血行转移模型,发现利用ISET外周血筛查装置的CTC的检出率为17.65%,显著低于利用ISET血液循环系统的检出率82.35%(P<0.05)。
结论：1.ISET技术在体外实验中有较高的敏感性和特异性,且实验方法简单,不依赖任何大型设备,实验结果稳定,可重复性强。2.在国际上首次利用ISET技术在肿瘤细胞血行转移动物模型中检测到CTC,具有极高的特异性。3.ISET外周血筛查装置适用于临床筛查CTC且方便组装便于携带。4.ISET技术在临床实验中表现出极高的特异性,但是敏感性不佳。5.ISET技术在疗效监视、评估预后以及在肿瘤转移机制研究方面有巨大潜力。6.在国际上首次设计并建立ISET血液循环系统,可以显著提高ISET技术敏感性,并且直接减少循环血液中存在的CTC数量。
上述技术、方法的实施对肿瘤诊断、疗效监视提供了新的思路。
OBJECTIVE:Systematic study of isolation by size of epithelial tumor cells (ISET) methodology in vitro, animal experiments and clinical trials. To establish a kind of method of ISET in the process of blood circulation to detect, screen and separate circulating tumor cells. This pilot study will serve as a basis for a much larger clinical trial.
METHOD:1.The method for the technical processes of ISET technology in vitro established with H520-pEGFP cells. H520-pEGFP cells will mix into the physiological saline and the blood sample from healthy volunteers respectively; detect to ascertain the recovery, the sensitivity and the specificity by ISET.2.To establish an animal model of tumor cells metastasis in the blood circulating, and detect the CTCs in the peripheral blood of animals by ISET technology.3.To design an ISET screening device, detect the CTCs in the peripheral blood of96patients with pulmonary disease. To observe the sensitivity and specificity of ISET screening device used in clinical testing. To monitor the presence of CTCs in one patient's peripheral circulation undergoing standard chemotherapy and radiotherapy.4.To design an ISET hemoperfusion system, detect the CTCs in the blood circulating of rabbit with VX2tumor, observe and compare sensitivity and security of ISET screening device and ISET hemoperfusion system.
RESULTS:1.The pEGFP-N3plasmid was transfected into H520tumor cells successfully. The recovery rate of mixed tumor cells detected by ISET technology remained about97.57±1.33in physiological saline and79.53±2.63in blood sample from healthy volunteers. We could detect1tumor cell in1ml peripheral blood while no positive cells were detected in healthy control, showing the high sensitivity and specificity of the ISET technology.2.The NOD-SCID mice model and rabbit model of tumor cells metastasis in the blood circulating have been established respectively.3samples in3NOD-SCID mice and3samples in17rabbit were detected with the CTCs by ISET technology.3.The ISET screening device was designed and built successfully to detect the CTCs in the peripheral blood of96patients with pulmonary disease.2samples in67cancer patients and0sample in29non-neoplastic patients were detected with the CTCs.2samples in4patients of stage IV and0sample in63patients of stage Ⅰ-Ⅲ were detected with the CTCs. However, CTCs were not detected in all6healthy samples by control. The detection rate of CTCs in4patients of stage IV was higher than that in63tumor patients of stage Ⅰ-Ⅲ (P<0.05).4.The persistence of CTCs in peripheral circulation in the patient of stage IV were monitored by ISET screening device.5.The ISET hemoperfusion system was designed and built successfully to detect the CTCs in the blood circulating of rabbit with VX2tumor. The detection rate of CTCs by ISET hemoperfusion system was higher than that by ISET screening device (P<0.05),82.35and17.65respectively (p<0.05).
Conclusion:1.The ISET technology to detect CTCs was convenient and stable with high sensitivity and good specificity. It does not rely on any large-scale experimental equipment.2.CTCs have been detected in animal models of tumor cells metastasis in the blood circulating at the first time in the international, indicating that ISET technology has a high specificity.3.ISET screening device suitable for clinical detecting CTCs and easy to assemble and carry.4.The ISET technology exhibit high specificity in clinical trials, but the sensitivity is poor.5.We can evaluate efficacy of chemotherapy and radiotherapy and other cancer-targeting therapies by using ISET technology that has great potential for mechanistic studies of ISET-isolated CTC biology.6.A new ISET hemoperfusion system could be designed to increase sensitivity to CTCs greatly and reduce the number of CTCs directly during the blood flow at the first time in the international.
The ISET hemoperfusion system gives us new insights into the diagnosis of malignant tumor and monitoring adjuvant therapy.
引文
[1]PANTEL K, ALIX-PANABIERES C. Circulating tumour cells in cancer patients: challenges and perspectives [J]. Trends in molecular medicine,2010,16(9): 398-406.
[2]PULTE D, LI E, CRAWFORD B K, et al. Sentinel lymph node mapping and molecular staging in nonsmall cell lung carcinoma [J]. Cancer,2005,104(7): 1453-61.
[3]IDDINGS D, BILCHIK A. The biologic significance of micrometastatic disease and sentinel lymph node technology on colorectal cancer [J]. Journal of surgical oncology,2007,96(8):671-7.
[4]BILCHIK A J, HOON D S, SAHA S, et al. Prognostic impact of micrometastases in colon cancer:interim results of a prospective multicenter trial [J]. Annals of surgery,2007,246(4):568-75; discussion 75-7.
[5]GRONAU E, GOPPELT M, HARZMANN R, et al. Prostate cancer relapse after therapy with curative intention:a diagnostic and therapeutic dilemma [J]. Onkologie,2005,28(6-7):361-6.
[6]LANG J E, HALL C S, SINGH B, et al. Significance of micrometastasis in bone marrow and blood of operable breast cancer patients:research tool or clinical application? [J]. Expert review of anticancer therapy,2007,7(10):1463-72.
[7]SWINDLE P W, KATTAN M W, SCARDINO P T. Markers and meaning of primary treatment failure [J]. The Urologic clinics of North America,2003,30(2): 377-401.
[8]PANTEL K, BRAKENHOFF R H. Dissecting the metastatic cascade [J]. Nature reviews Cancer,2004,4(6):448-56.
[9]WIKMAN H, VESSELLA R, PANTEL K. Cancer micrometastasis and tumour dormancy [J]. APMIS:acta pathologica, microbiologica, et immunologica Scandinavica,2008,116(7-8):754-70.
[10]RIETHDORF S, WIKMAN H, PANTEL K. Review:Biological relevance of disseminated tumor cells in cancer patients [J]. International journal of cancer Journal international du cancer,2008,123(9):1991-2006.
[11]ROSENBERG R, GERTLER R, FRIEDERICHS J, et al. Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood [J]. Cytometry,2002,49(4):150-8.
[12]VONA G, SABILE A, LOUHA M, et al. Isolation by size of epithelial tumor cells:a new method for the immunomorphological and molecular characterization of circulatingtumor cells [J]. The American journal of pathology, 2000,156(1):57-63.
[13]PANTEL K, BRAKENHOFF R H, BRANDT B. Detection, clinical relevance and specific biological properties of disseminating tumour cells [J]. Nature reviews Cancer,2008,8(5):329-40.
[14]LU J, FAN T, ZHAO Q, et al. Isolation of circulating epithelial and tumor progenitor cells with an invasive phenotype from breast cancer patients [J]. International journal of cancer Journal international du cancer,2010,126(3): 669-83.
[15]GAFORIO J J, SERRANO M J, SANCHEZ-ROVIRA P, et al. Detection of breast cancer cells in the peripheral blood is positively correlated with estrogen-receptor status and predicts for poor prognosis [J]. International journal of cancer Journal international du cancer,2003,107(6):984-90.
[16]WULFING P, BORCHARD J, BUERGER H, et al. HER2-positive circulating tumor cells indicate poor clinical outcome in stage I to III breast cancer patients [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(6):1715-20.
[17]SIENEL W, SEEN-HIBLER R, MUTSCHLER W, et al. Tumour cells in the tumour draining vein of patients with non-small cell lung cancer:detection rate and clinical significance [J]. European journal of cardio-thoracic surgery: official journal of the European Association for Cardio-thoracic Surgery,2003, 23(4):451-6.
[18]PANTEL K, ALIX-PANABIERES C, RIETHDORF S. Cancer micrometastases [J]. Nature reviews Clinical oncology,2009,6(6):339-51.
[19]HAYES D F, CRISTOFANILLI M, BUDD G T, et al. Circulating tumor cells at each follow-up time point during therapy of metastatic breast cancer patients predict progression-free and overall survival [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(14 Pt 1):4218-24.
[20]BIDARD F C, VINCENT-SALOMON A, SIGAL-ZAFRANI B, et al. Prognosis of women with stage IV breast cancer depends on detection of circulating tumor cells rather than disseminated tumor cells [J]. Annals of oncology:official journal of the European Society for Medical Oncology/ESMO,2008,19(3): 496-500.
[21]STOTT S L, LEE R J, NAGRATH S, et al. Isolation and characterization of circulating tumor cells from patients with localized and metastatic prostate cancer [J]. Science translational medicine,2010,2(25):25ra3.
[22]MAHESWARAN S, SEQUIST L V, NAGRATH S, et al. Detection of mutations in EGFR in circulating lung-cancer cells [J]. The New England journal of medicine,2008,359(4):366-77.
[23]NAGRATH S, SEQUIST L V, MAHESWARAN S, et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology [J]. Nature, 2007,450(7173):1235-9.
[24]HELZER K T, BARNES H E, DAY L, et al. Circulating tumor cells are transcriptionally similar to the primary tumor in a murine prostate model [J]. Cancer research,2009,69(19):7860-6.
[25]VREDENBURGH J J, SILVA O, TYER C, et al. A comparison of immunohistochemistry, two-color immunofluorescence, and flow cytometry with cell sorting for the detection of micrometastatic breast cancer in the bone marrow [J]. Journal of hematotherapy,1996,5(1):57-62.
[26]ALIX-PANABIERES C, VENDRELL J P, SLIJPER M, et al. Full-length cytokeratin-19 is released by human tumor cells:a potential role in metastatic progression of breast cancer [J]. Breast cancer research:BCR,2009,11(3):R39.
[27]SCHWARZENBACH H, ALIX-PANABIERES C, MULLER I, et al. Cell-free tumor DNA in blood plasma as a marker for circulating tumor cells in prostate cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2009,15(3):1032-8.
[28]APOSTOLOPOULOS C, GIAMAS G, STEBBING J. Clinical significance of circulating tumor cells [J]. Biomarkers in medicine,2007,1(3):341-2.
[29]ALIX-PANABIERES C, VENDRELL J P, PELLE O, et al. Detection and characterization of putative metastatic precursor cells in cancer patients [J]. Clinical chemistry,2007,53(3):537-9.
[30]ALIX-PANABIERES C, REBILLARD X, BROUILLET J P, et al. Detection of circulating prostate-specific antigen-secreting cells in prostate cancer patients [J]. Clinical chemistry,2005,51(8):1538-41.
[31]PACHMANN K, CAMARA O, KAVALLARIS A, et al. Monitoring the response of circulating epithelial tumor cells to adjuvant chemotherapy in breast cancer allows detection of patients at risk of early relapse [J]. Journal of clinical oncology:official journal of the American Society of Clinical Oncology,2008, 26(8):1208-15.
[32]PACHMANN K, CLEMENT J H, SCHNEIDER C P, et al. Standardized quantification of circulating peripheral tumor cells from lung and breast cancer [J]. Clinical chemistry and laboratory medicine:CCLM/FESCC,2005,43(6): 617-27.
[33]YAMASHITA J, MATSUO A, KURUSU Y, et al. Preoperative evidence of circulating tumor cells by means of reverse transcriptase-polymerase chain reaction for carcinoembryonic antigen messenger RNA is an independent predictor of survival in non-small cell lung cancer:a prospective study [J]. The Journal of thoracic and cardiovascular surgery,2002,124(2):299-305.
[34]SHER Y P, SHIH J Y, YANG P C, et al. Prognosis of non-small cell lung cancer patients by detecting circulating cancer cells in the peripheral blood with multiple marker genes [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2005,11(1):173-9.
[35]CHEN T F, JIANG G L, FU X L, et al. CK19 mRNA expression measured by reverse-transcription polymerase chain reaction (RT-PCR) in the peripheral blood of patients with non-small cell lung cancer treated by chemo-radiation:an independent prognostic factor [J]. Lung Cancer,2007,56(1):105-14.
[36]MENG S, TRIPATHY D, FRENKEL E P, et al. Circulating tumor cells in patients with breast cancer dormancy [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2004,10(24): 8152-62.
[37]EDERER F, GOLDBLATT S A, NADEL E M. Analysis of the Color Micrograph Study of the Circulating Cancer Cell Cooperative (Cccc) [J]. Acta cytologica, 1965,9(50-7.
[38]LIU L, LIAO G Q, HE P, et al. Detection of circulating cancer cells in lung cancer patients with a panel of marker genes [J]. Biochemical and biophysical research communications,2008,372(4):756-60.
[39]SHEU C C, CHANG M Y, CHANG H C, et al. Combined detection of CEA, CK-19 and c-met mRNAs in peripheral blood:a highly sensitive panel for potential molecular diagnosis of non-small cell lung cancer [J]. Oncology,2006, 70(3):203-11.
[40]YAMAMOTO O, TAKAHASHI H, HIRASAWA M, et al. Surfactant protein gene expressions for detection of lung carcinoma cells in peripheral blood [J]. Respiratory medicine,2005,99(9):1164-74.
[41]ROSS A A, COOPER B W, LAZARUS H M, et al. Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques [J]. Blood,1993, 82(9):2605-10.
[42]MOSTERT B, SLEIJFER S, FOEKENS J A, et al. Circulating tumor cells (CTCs):detection methods and their clinical relevance in breast cancer [J]. Cancer treatment reviews,2009,35(5):463-74.
[43]SHIMOMURA O. Discovery of green fluorescent protein (GFP) (Nobel Lecture) [J]. Angew Chem Int Ed Engl,2009,48(31):5590-602.
[44]CORMACK B P, VALDIVIA R H, FALKOW S. FACS-optimized mutants of the green fluorescent protein (GFP) [J]. Gene,1996,173(1 Spec No):33-8.
[45]ZABAGLO L, ORMEROD M G, PARTON M, et al. Cell filtration-laser scanning cytometry for the characterisation of circulating breast cancer cells [J]. Cytometry Part A:the journal of the International Society for Analytical Cytology,2003,55(2):102-8.
[46]DE GRAAFF M A, CLETON-JANSEN A M, SZUHAI K, et al. Mediator complex subunit 12 exon 2 mutation analysis in different subtypes of smooth muscle tumors confirms genetic heterogeneity [J]. Human pathology,2013,
[47]WANG Z A, MITROFANOVA A, BERGREN S K, et al. Lineage analysis of basal epithelial cells reveals their unexpected plasticity and supports a cell-of-origin model for prostate cancer heterogeneity [J]. Nature cell biology, 2013,15(3):274-83.
[48]MANI S A, GUO W, LIAO M J, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells [J]. Cell,2008,133(4):704-15.
[49]BERX G, RASPE E, CHRISTOFORI G, et al. Pre-EMTing metastasis? Recapitulation of morphogenetic processes in cancer [J]. Clinical & experimental metastasis,2007,24(8):587-97.
[50]CHENG W Y, KANDEL J J, YAMASHIRO D J, et al. A multi-cancer mesenchymal transition gene expression signature is associated with prolonged time to recurrence in glioblastoma [J]. PloS one,2012,7(4):e34705.
[51]STOYIANNI A, GOUSSIA A, PENTHEROUDAKIS G, et al. Immunohistochemical study of the epithelial-mesenchymal transition phenotype in cancer of unknown primary:incidence, correlations and prognostic utility [J]. Anticancer research,2012,32(4):1273-81.
[52]SUGIMOTO H, ITO M, HATANO M, et al. Roles of epithelial-mesenchymal transition in squamous cell carcinoma of the temporal bone [J]. Otology & neurotology:official publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology, 2011,32(3):483-7.
[53]PINZANI P, SALVADORI B, SIMI L, et al. Isolation by size of epithelial tumor cells in peripheral blood of patients with breast cancer:correlation with real-time reverse transcriptase-polymerase chain reaction results and feasibility of molecular analysis by laser microdissection [J]. Human pathology,2006,37(6): 711-8.
[54]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Application of a filtration- and isolation-by-size technique for the detection of circulating tumor cells in cutaneous melanoma [J]. The Journal of investigative dermatology,2010, 130(10):2440-7.
[55]HOFMAN V, ILIE M I, LONG E, et al. Detection of circulating tumor cells as a prognostic factor in patients undergoing radical surgery for non-small-cell lung carcinoma:comparison of the efficacy of the CellSearch Assay and the isolation by size of epithelial tumor cell method [J]. International journal of cancer Journal international du cancer,2011,129(7):1651-60.
[56]HOFMAN V, BONNETAUD C, ILIE M I, et al. Preoperative circulating tumor cell detection using the isolation by size of epithelial tumor cell method for patients with lung cancer is a new prognostic biomarker [J]. Clinical cancer research:an official journal of the American Association for Cancer Research, 2011,17(4):827-35.
[57]HOFMAN V J, ILIE M I, BONNETAUD C, et al. Cytopathologic detection of circulating tumor cells using the isolation by size of epithelial tumor cell method: promises and pitfalls [J]. American journal of clinical pathology,2011,135(1): 146-56.
[58]KIDD J G, ROUS P. Cancers Deriving from the Virus Papillomas of Wild Rabbils under Natural Conditions [J]. The Journal of experimental medicine, 1940,71(4):469-94.
[59]GIRI I, DANOS O, YANIV M. Genomic structure of the cottontail rabbit (Shope) papillomavirus [J]. Proceedings of the National Academy of Sciences of the United States of America,1985,82(6):1580-4.
[60]WOYWODT A, GOLDBERG C, SCHEER J, et al. An improved assay for enumeration of circulating endothelial cells [J]. Annals of hematology,2004, 83(8):491-4.
[61]KREBS M G., HOU J M, SLOANE R, et al. Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and-independent approaches [J]. Journal of thoracic oncology:official publication of the International Association for the Study of Lung Cancer,2012,7(2): 306-15.
[62]WERLING R W, YAZIJI H, BACCHI C E, et al. CDX2, a highly sensitive and specific marker of adenocarcinomas of intestinal origin:an immunohistochemical survey of 476 primary and metastatic carcinomas [J]. The American journal of surgical pathology,2003,27(3):303-10.
[63]BOCKHORN M, JAIN R K, MUNN L L. Active versus passive mechanisms in metastasis:do cancer cells crawl into vessels, or are they pushed? [J]. The lancet oncology,2007,8(5):444-8.
[64]CRISTOFANILLI M, BUDD G T, ELLIS M J, et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer [J]. The New England journal of medicine,2004,351(8):781-91.
[65]BUDD G T, CRISTOFANILLI M, ELLIS M J, et al. Circulating tumor cells versus imaging--Predicting overall survival in metastatic breast cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(21):6403-9.
[66]PIERGA J Y, BIDARD F C, MATHIOT C, et al. Circulating tumor cell detection predicts early metastatic relapse after neoadjuvant chemotherapy in large operable and locally advanced breast cancer in a phase II randomized trial [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2008,14(21):7004-10.
[67]PESTRIN M, BESSIS, GALARDI F, et al. Correlation of HER2 status between primary tumors and corresponding circulating tumor cells in advanced breast cancer patients [J]. Breast cancer research and treatment,2009,118(3):523-30.
[68]KALLURI R, WEINBERG R A. The basics of epithelial-mesenchymal transition [J]. The Journal of clinical investigation,2009,119(6):1420-8.
[69]THIERY J P. Epithelial-mesenchymal transitions in tumour progression [J]. Nature reviews Cancer,2002,2(6):442-54.
[70]THIERY J P, ACLOQUE H, HUANG R Y, et al. Epithelial-mesenchymal transitions in development and disease [J]. Cell,2009,139(5):871-90.
[71]MEGO M, MANI S A, CRISTOFANILLI M. Molecular mechanisms of metastasis in breast cancer--clinical applications [J]. Nature reviews Clinical oncology,2010,7(12):693-701.
[72]ALLARD W J, MATERA J, MILLER M C, et al. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2004,10(20):6897-904.
[73]FARACE F, MASSARD C, VIMOND N, et al. A direct comparison of CellSearch and ISET for circulating tumour-cell detection in patients with metastatic carcinomas [J]. British journal of cancer,2011,105(6):847-53.
[74]KHOJA L, BACKEN A, SLOANE R, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker [J]. British journal of cancer,2012,106(3):508-16.
[75]LIOTTA L A, KLEINERMAN J, SAIDEL G M. Quantitative relationships of intravascular tumor cells, tumor vessels, and pulmonary metastases following tumor implantation [J]. Cancer research,1974,34(5):997-1004.
[76]WEITZ J, KIENLE P, LACROIX J, et al. Dissemination of tumor cells in patients undergoing surgery for colorectal cancer [J]. Clinical cancer research: an official journal of the American Association for Cancer Research,1998,4(2): 343-8.
[77]ESCHWEGE P, MOUTEREAU S, DROUPY S, et al. Prognostic value of prostate circulating cells detection in prostate cancer patients:a prospective study [J]. British journal of cancer,2009,100(4):608-10.
[78]SCHMIDT T, KOCH M, ANTOLOVIC D, et al. Influence of two different resection techniques (conventional liver resection versus anterior approach) of liver metastases from colorectal cancer on hematogenous tumor cell dissemination-prospective randomized multicenter trial [J]. BMC surgery,2008, 8(6.
[79]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Circulating benign nevus cells detected by ISET technique:warning for melanoma molecular diagnosis [J]. Archives of dermatology,2010,146(10):1120-4.
[1]WORLD HEALTH ORGANIZATION. The global burden of disease:2004 update [M]. Geneva: World Health Organization,2008.
[2]KOHN E C, LIOTTA L A. Molecular insights into cancer invasion:strategies for prevention and intervention [J]. Cancer research,1995,55(9):1856-62.
[3]FIDLER I J. Tumor heterogeneity and the biology of cancer invasion and metastasis [J]. Cancer research,1978,38(9):2651-60.
[4]GUPTA G P, MASS AGUE J. Cancer metastasis: building a framework [J]. Cell, 2006,127(4):679-95.
[5]MOSTERT B, SLEIJFER S, FOEKENS J A, et al. Circulating tumor cells (CTCs): detection methods and their clinical relevance in breast cancer [J]. Cancer treatment reviews,2009,35(5):463-74.
[6]ROSS A A, COOPER B W, LAZARUS H M, et al. Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques [J]. Blood,1993, 82(9):2605-10.
[7]VONA G, SABILE A, LOUHA M, et al. Isolation by size of epithelial tumor cells: a new method for the immunomorphological and molecular characterization of circulatingtumor cells [J]. The American journal of pathology,2000,156(1): 57-63.
[8]MENG S, TRIPATHY D, FRENKEL E P, et al. Circulating tumor cells in patients with breast cancer dormancy [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2004,10(24):8152-62.
[9]ZABAGLO L, ORMEROD M G, PARTON M, et al. Cell filtration-laser scanning cytometry for the characterisation of circulating breast cancer cells [J]. Cytometry Part A:the journal of the International Society for Analytical Cytology,2003,55(2):102-8.
[10]PATERLINI-BRECHOT P, BENALI N L. Circulating tumor cells (CTC) detection: clinical impact and future directions [J]. Cancer letters,2007,253(2): 180-204.
[11]WOYWODT A, GOLDBERG C, SCHEER J, et al. An improved assay for enumeration of circulating endothelial cells [J]. Annals of hematology,2004, 83(8):491-4.
[12]HOFMAN V, BONNETAUD C, ILIE M I, et al. Preoperative circulating tumor cell detection using the isolation by size of epithelial tumor cell method for patients with lung cancer is a new prognostic biomarker [J]. Clinical cancer research:an official journal of the American Association for Cancer Research, 2011,17(4):827-35.
[13]HOFMAN V J, ILIE M I, BONNETAUD C, et al. Cytopathologic detection of circulating tumor cells using the isolation by size of epithelial tumor cell method: promises and pitfalls [J]. American journal of clinical pathology,2011,135(1): 146-56.
[14]KREBS M G, HOU J M, SLOANE R, et al. Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and-independent approaches [J]. Journal of thoracic oncology:official publication of the International Association for the Study of Lung Cancer,2012,7(2): 306-15.
[15]HRISTOZOVA T, KONSCHAK R, BUDACH V, et al. A simple multicolor flow cytometry protocol for detection and molecular characterization of circulating tumor cells in epithelial cancers [J]. Cytometry Part A:the journal of the International Society for Analytical Cytology,2012,
[16]COHEN S J, ALPAUGH R K, GROSS S, et al. Isolation and characterization of circulating tumor cells in patients with metastatic colorectal cancer [J]. Clinical colorectal cancer,2006,6(2):125-32.
[17]STRIJBOS M H, GRATAMA J W, KRAAN J, et al. Circulating endothelial cells in oncology: pitfalls and promises [J]. British journal of cancer,2008,98(11): 1731-5.
[18]SMIRNOV D A, FOULK B W, DOYLE G V, et al. Global gene expression profiling of circulating endothelial cells in patients with metastatic carcinomas [J]. Cancer research,2006,66(6):2918-22.
[19]HOU J M, KREBS M, WARD T, et al. Circulating tumor cells as a window on metastasis biology in lung cancer [J]. The American journal of pathology,2011, 178(3):989-96.
[20]KHOJA L, BACKEN A, SLOANE R, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker [J]. British journal of cancer,2012,106(3):508-16.
[21]VONA G, ESTEPA L, BEROUD C, et al. Impact of cytomorphological detection of circulating tumor cells in patients with liver cancer [J]. Hepatology, 2004,39(3):792-7.
[22]PINZANI P, SALVADORI B, SIMI L, et al. Isolation by size of epithelial tumor cells in peripheral blood of patients with breast cancer: correlation with real-time reverse transcriptase-polymerase chain reaction results and feasibility of molecular analysis by laser microdissection [J]. Human pathology,2006,37(6): 711-8.
[23]PANTEL K, ALIX-PANABIERES C, RIETHDORF S. Cancer micrometastases [J]. Nature reviews Clinical oncology,2009,6(6):339-51.
[24]HAYES D F, CRISTOFANILLI M, BUDD G T, et al. Circulating tumor cells at each follow-up time point during therapy of metastatic breast cancer patients predict progression-free and overall survival [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(14 Pt 1):4218-24.
[25]BIDARD F C, VINCENT-SALOMON A, SIGAL-ZAFRANI B, et al. Prognosis of women with stage IV breast cancer depends on detection of circulating tumor cells rather than disseminated tumor cells [J]. Annals of oncology:official journal of the European Society for Medical Oncology/ESMO,2008,19(3): 496-500.
[26]STOTT S L, LEE R J, NAGRATH S, et al. Isolation and characterization of circulating tumor cells from patients with localized and metastatic prostate cancer [J]. Science translational medicine,2010,2(25):25ra3.
[27]MAHESWARAN S, SEQUIST L V, NAGRATH S, et al. Detection of mutations in EGFR in circulating lung-cancer cells [J]. The New England journal of medicine,2008,359(4):366-77.
[28]NAGRATH S, SEQUIST L V, MAHESWARAN S, et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology [J]. Nature, 2007,450(7173):1235-9.
[29]HELZER K T, BARNES H E, DAY L, et al. Circulating tumor cells are transcriptionally similar to the primary tumor in a murine prostate model [J]. Cancer research,2009,69(19):7860-6.
[30]VREDENBURGH J J, SILVA O, TYER C, et al. A comparison of immunohistochemistry, two-color immunofluorescence, and flow cytometry with cell sorting for the detection of micrometastatic breast cancer in the bone marrow [J]. Journal of hematotherapy,1996,5(1):57-62.
[31]ALIX-PANABIERES C, VENDRELL J P, SLIJPER M, et al. Full-length cytokeratin-19 is released by human tumor cells:a potential role in metastatic progression of breast cancer [J]. Breast cancer research:BCR,2009,11(3):R39.
[32]SCHWARZENBACH H, ALIX-PANABIERES C, MULLER I, et al. Cell-free tumor DNA in blood plasma as a marker for circulating tumor cells in prostate cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2009,15(3):1032-8.
[33]APOSTOLOPOULOS C, GIAMAS G, STEBBING J. Clinical significance of circulating tumor cells [J]. Biomarkers in medicine,2007,1(3):341-2.
[34]ALIX-PANABIERES C, VENDRELL J P, PELLE O, et al. Detection and characterization of putative metastatic precursor cells in cancer patients [J]. Clinical chemistry,2007,53(3):537-9.
[35]ALIX-PANABIERES C, REBILLARD X, BROUILLET J P, et al. Detection of circulating prostate-specific antigen-secreting cells in prostate cancer patients [J]. Clinical chemistry,2005,51(8):1538-41.
[36]PACHMANN K, CAMARA O, KAVALLARIS A, et al. Monitoring the response of circulating epithelial tumor cells to adjuvant chemotherapy in breast cancer allows detection of patients at risk of early relapse [J]. Journal of clinical oncology:official journal of the American Society of Clinical Oncology,2008, 26(8):1208-15.
[37]PACHMANN K, CLEMENT J H, SCHNEIDER C P, et al. Standardized quantification of circulating peripheral tumor cells from lung and breast cancer [J]. Clinical chemistry and laboratory medicine:CCLM/FESCC,2005,43(6): 617-27.
[38]LU J, FAN T, ZHAO Q, et al. Isolation of circulating epithelial and tumor progenitor cells with an invasive phenotype from breast cancer patients [J]. International journal of cancer Journal international du cancer,2010,126(3): 669-83.
[39]PANTEL K, BRAKENHOFF R H, BRANDT B. Detection, clinical relevance and specific biological properties of disseminating tumour cells [J]. Nature reviews Cancer,2008,8(5):329-40.
[40]BENOY I H, ELST H, PHILIPS M, et al. Real-time RT-PCR detection of disseminated tumour cells in bone marrow has superior prognostic significance in comparison with circulating tumour cells in patients with breast cancer [J]. British journal of cancer,2006,94(5):672-80.
[41]SCHMITT M, FOEKENS J A. Circulating tumor cells in blood of primary breast cancer patients assessed by a novel RT-PCR test kit and comparison with status of bone marrow-disseminated tumor cells [J]. Breast cancer research:BCR, 2009,11(5):109.
[42]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Application of a filtration-and isolation-by-size technique for the detection of circulating tumor cells in cutaneous melanoma [J]. The Journal of investigative dermatology,2010, 130(10):2440-7.
[43]HOFMAN V, ILIE M I, LONG E, et al. Detection of circulating tumor cells as a prognostic factor in patients undergoing radical surgery for non-small-cell lung carcinoma:comparison of the efficacy of the CellSearch Assay and the isolation by size of epithelial tumor cell method [J]. International journal of cancer Journal international du cancer,2011,129(7):1651-60.
[44]DE ALBUQUERQUE A, KUBISCH I, BREIER G, et al. Multimarker gene analysis of circulating tumor cells in pancreatic cancer patients:a feasibility study [J]. Oncology,2012,82(1):3-10.
[45]SALOUSTROS E, PERRAKI M, APOSTOLAKI S, et al. Cytokeratin-19 mRNA-positive circulating tumor cells during follow-up of patients with operable breast cancer:prognostic relevance for late relapse [J]. Breast cancer research:BCR,2011,13(3):R60.
[46]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Circulating benign nevus cells detected by ISET technique:warning for melanoma molecular diagnosis [J]. Archives of dermatology,2010,146(10):1120-4.
[47]SCHUSTER R, BECHRAKIS N E, STROUX A, et al. Prognostic relevance of circulating tumor cells in metastatic uveal melanoma [J]. Oncology,2011, 80(1-2):57-62.
[48]PINZANI P, MAZZINI C, SALVIANTI F, et al. Tyrosinase mRNA levels in the blood of uveal melanoma patients:correlation with the number of circulating tumor cells and tumor progression [J]. Melanoma research,2010,20(4):303-10.
[49]PECK K, SHER Y P, SHIH J Y, et al. Detection and quantitation of circulating cancer cells in the peripheral blood of lung cancer patients [J]. Cancer research, 1998,58(13):2761-5.
[50]PANTELEAKOU Z, LEMBESSIS P, SOURLA A, et al. Detection of circulating tumor cells in prostate cancer patients:methodological pitfalls and clinical relevance [J]. Mol Med,2009,15(3-4):101-14.
[51]KUTUN S, CELIK A, CEM KOCKAR M, et al. Expression of CK-19 and CEA mRNA in peripheral blood of gastric cancer patients [J]. Experimental oncology, 2010,32(4):263-8.
[52]LANKIEWICZ S, ROTHER E, ZIMMERMANN S, et al. Tumour-associated transcripts and EGFR deletion variants in colorectal cancer in primary tumour, metastases and circulating tumour cells [J]. Cellular oncology:the official journal of the International Society for Cellular Oncology,2008,30(6):463-71.
[53]BECKER S, BECKER-PERGOLA G, FEHM T, et al. Time is an important factor when processing samples for the detection of disseminated tumor cells in blood/bone marrow by reverse transcription-PCR [J]. Clinical chemistry,2004, 50(4):785-6.
[54]SASTRE J, MAESTRO M L, PUENTE J, et al. Circulating tumor cells in colorectal cancer:correlation with clinical and pathological variables [J]. Annals of oncology:official journal of the European Society for Medical Oncology/ ESMO,2008,19(5):935-8.
[55]DE BONO J S, SCHER H I, MONTGOMERY R B, et al. Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2008,14(19):6302-9.
[56]RIETHDORF S, FRITSCHE H, MULLER V, et al. Detection of circulating tumor cells in peripheral blood of patients with metastatic breast cancer:a validation study of the CellSearch system [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2007,13(3): 920-8.
[57]FARACE F, MASSARD C, VIMOND N, et al. A direct comparison of CellSearch and ISET for circulating tumour-cell detection in patients with metastatic carcinomas [J]. British journal of cancer,2011,105(6):847-53.
[58]MEDINA A, BROWN E, CARR N, et al. Circulating monocytes have the capacity to be transdifferentiated into keratinocyte-like cells [J]. Wound repair and regeneration:official publication of the Wound Healing Society [and] the European Tissue Repair Society,2009,17(2):268-77.
[59]CHENG W Y, KANDEL J J, YAMASHIRO D J, et al. A multi-cancer mesenchymal transition gene expression signature is associated with prolonged time to recurrence in glioblastoma [J]. PloS one,2012,7(4):e34705.
[60]STOYIANNI A, GOUSSIA A, PENTHEROUDAKIS G, et al. Immunohistochemical study of the epithelial-mesenchymal transition phenotype in cancer of unknown primary:incidence, correlations and prognostic utility [J]. Anticancer research,2012,32(4):1273-81.
[61]SUGIMOTO H, ITO M, HATANO M, et al. Roles of epithelial-mesenchymal transition in squamous cell carcinoma of the temporal bone [J]. Otology & neurotology:official publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology, 2011,32(3):483-7.
[62]BERX G, RASPE E, CHRISTOFORI G, et al. Pre-EMTing metastasis? Recapitulation of morphogenetic processes in cancer [J]. Clinical & experimental metastasis,2007,24(8):587-97.
[63]KALLURI R, WEINBERG R A. The basics of epithelial-mesenchymal transition [J]. The Journal of clinical investigation,2009,119(6):1420-8.
[64]THIERY J P. Epithelial-mesenchymal transitions in tumour progression [J]. Nature reviews Cancer,2002,2(6):442-54.
[65]MANI S A, GUO W, LIAO M J, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells [J]. Cell,2008,133(4):704-15.
[66]POLYAK K, WEINBERG R A. Transitions between epithelial and mesenchymal states:acquisition of malignant and stem cell traits [J]. Nature reviews Cancer,2009,9(4):265-73.
[67]THIERY J P, ACLOQUE H, HUANG R Y, et al. Epithelial-mesenchymal transitions in development and disease [J]. Cell,2009,139(5):871-90.
[68]LECHARPENTIER A, VIELH P, PEREZ-MORENO P, et al. Detection of circulating tumour cells with a hybrid (epithelial/mesenchymal) phenotype in patients with metastatic non-small cell lung cancer [J]. British journal of cancer, 2011,105(9):1338-41.
[69]OZAKI K, YAMAGAMI T, NOMURA K, et al. Prognostic significance of surgical margin, Ki-67 and cyclin D1 protein expression in grade II canine cutaneous mast cell tumor [J]. The Journal of veterinary medical science/the Japanese Society of Veterinary Science,2007,69(11):1117-21.
[70]BALIC M, LIN H, YOUNG L, et al. Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(19):5615-21.
[71]ALLAN A L, VANTYGHEM S A, TUCK A B, et al. Tumor dormancy and cancer stem cells:implications for the biology and treatment of breast cancer metastasis [J]. Breast disease,2006,26(87-98.
[72]HOFMAN V, LONG E, ILIE M, et al. Morphological analysis of circulating tumour cells in patients undergoing surgery for non-small cell lung carcinoma using the isolation by size of epithelial tumour cell (ISET) method [J]. Cytopathology:official journal of the British Society for Clinical Cytology, 2012,23(1):30-8.
[73]MAHESWARAN S, HABER D A. Circulating tumor cells:a window into cancer biology and metastasis [J]. Current opinion in genetics & development, 2010,20(1):96-9.
[74]LELIEVRE L, PATERLINI-BRECHOT P, CAMATTE S, et al. Effect of laparoscopy versus laparotomy on circulating tumor cells using isolation by size of epithelial tumor cells [J]. International journal of gynecological cancer official journal of the International Gynecological Cancer Society,2004,14(2): 229-33.
[75]NANAL R, KYLE P, SOOTHILL P W. A classification of pregnancy losses after invasive prenatal diagnostic procedures:an approach to allow comparison of units with a different case mix [J]. Prenatal diagnosis,2003,23(6):488-92.
[76]BIANCHI D W. Fetal cells in the maternal circulation:feasibility for prenatal diagnosis [J]. British journal of haematology,1999,105(3):574-83.
[77]NICE. Antenatal care-routine care for the healthy pregnant woman-clinical guideline [M]. London:RCOG Press (NICE Clinical Guidelines, No.62.).2008 Mar.
[78]FINNING K M, MARTIN P G, SOOTHILL P W, et al. Prediction of fetal D status from maternal plasma:introduction of a new noninvasive fetal RHD genotyping service [J]. Transfusion,2002,42(8):1079-85.
[79]VONA G, BEROUD C, BENACHI A, et al. Enrichment, immunomorphological, and genetic characterization of fetal cells circulating in maternal blood [J]. The American journal of pathology,2002,160(1):51-8.
[80]BEROUD C, KARLIOVA M, BONNEFONT J P, et al. Prenatal diagnosis of spinal muscular atrophy by genetic analysis of circulating fetal cells [J]. Lancet, 2003,361(9362):1013-4.
[81]SAKER A, BENACHI A, BONNEFONT J P, et al. Genetic characterisation of circulating fetal cells allows non-invasive prenatal diagnosis of cystic fibrosis [J]. Prenatal diagnosis,2006,26(10):906-16.
[82]FUNAKI S, SAWABATA N, NAKAGIRI T, et al. Novel approach for detection of isolated tumor cells in pulmonary vein using negative selection method: morphological classification and clinical implications [J]. European journal of cardio-thoracic surgery:official journal of the European Association for Cardio-thoracic Surgery,2011,40(2):322-7.
[83]MEHES G, WITT A, KUBISTA E, et al. Circulating breast cancer cells are frequently apoptotic [J]. The American journal of pathology,2001,159(1):17-20.
[2]PULTE D, LI E, CRAWFORD B K, et al. Sentinel lymph node mapping and molecular staging in nonsmall cell lung carcinoma [J]. Cancer,2005,104(7): 1453-61.
[3]IDDINGS D, BILCHIK A. The biologic significance of micrometastatic disease and sentinel lymph node technology on colorectal cancer [J]. Journal of surgical oncology,2007,96(8):671-7.
[4]BILCHIK A J, HOON D S, SAHA S, et al. Prognostic impact of micrometastases in colon cancer:interim results of a prospective multicenter trial [J]. Annals of surgery,2007,246(4):568-75; discussion 75-7.
[5]GRONAU E, GOPPELT M, HARZMANN R, et al. Prostate cancer relapse after therapy with curative intention:a diagnostic and therapeutic dilemma [J]. Onkologie,2005,28(6-7):361-6.
[6]LANG J E, HALL C S, SINGH B, et al. Significance of micrometastasis in bone marrow and blood of operable breast cancer patients:research tool or clinical application? [J]. Expert review of anticancer therapy,2007,7(10):1463-72.
[7]SWINDLE P W, KATTAN M W, SCARDINO P T. Markers and meaning of primary treatment failure [J]. The Urologic clinics of North America,2003,30(2): 377-401.
[8]PANTEL K, BRAKENHOFF R H. Dissecting the metastatic cascade [J]. Nature reviews Cancer,2004,4(6):448-56.
[9]WIKMAN H, VESSELLA R, PANTEL K. Cancer micrometastasis and tumour dormancy [J]. APMIS:acta pathologica, microbiologica, et immunologica Scandinavica,2008,116(7-8):754-70.
[10]RIETHDORF S, WIKMAN H, PANTEL K. Review:Biological relevance of disseminated tumor cells in cancer patients [J]. International journal of cancer Journal international du cancer,2008,123(9):1991-2006.
[11]ROSENBERG R, GERTLER R, FRIEDERICHS J, et al. Comparison of two density gradient centrifugation systems for the enrichment of disseminated tumor cells in blood [J]. Cytometry,2002,49(4):150-8.
[12]VONA G, SABILE A, LOUHA M, et al. Isolation by size of epithelial tumor cells:a new method for the immunomorphological and molecular characterization of circulatingtumor cells [J]. The American journal of pathology, 2000,156(1):57-63.
[13]PANTEL K, BRAKENHOFF R H, BRANDT B. Detection, clinical relevance and specific biological properties of disseminating tumour cells [J]. Nature reviews Cancer,2008,8(5):329-40.
[14]LU J, FAN T, ZHAO Q, et al. Isolation of circulating epithelial and tumor progenitor cells with an invasive phenotype from breast cancer patients [J]. International journal of cancer Journal international du cancer,2010,126(3): 669-83.
[15]GAFORIO J J, SERRANO M J, SANCHEZ-ROVIRA P, et al. Detection of breast cancer cells in the peripheral blood is positively correlated with estrogen-receptor status and predicts for poor prognosis [J]. International journal of cancer Journal international du cancer,2003,107(6):984-90.
[16]WULFING P, BORCHARD J, BUERGER H, et al. HER2-positive circulating tumor cells indicate poor clinical outcome in stage I to III breast cancer patients [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(6):1715-20.
[17]SIENEL W, SEEN-HIBLER R, MUTSCHLER W, et al. Tumour cells in the tumour draining vein of patients with non-small cell lung cancer:detection rate and clinical significance [J]. European journal of cardio-thoracic surgery: official journal of the European Association for Cardio-thoracic Surgery,2003, 23(4):451-6.
[18]PANTEL K, ALIX-PANABIERES C, RIETHDORF S. Cancer micrometastases [J]. Nature reviews Clinical oncology,2009,6(6):339-51.
[19]HAYES D F, CRISTOFANILLI M, BUDD G T, et al. Circulating tumor cells at each follow-up time point during therapy of metastatic breast cancer patients predict progression-free and overall survival [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(14 Pt 1):4218-24.
[20]BIDARD F C, VINCENT-SALOMON A, SIGAL-ZAFRANI B, et al. Prognosis of women with stage IV breast cancer depends on detection of circulating tumor cells rather than disseminated tumor cells [J]. Annals of oncology:official journal of the European Society for Medical Oncology/ESMO,2008,19(3): 496-500.
[21]STOTT S L, LEE R J, NAGRATH S, et al. Isolation and characterization of circulating tumor cells from patients with localized and metastatic prostate cancer [J]. Science translational medicine,2010,2(25):25ra3.
[22]MAHESWARAN S, SEQUIST L V, NAGRATH S, et al. Detection of mutations in EGFR in circulating lung-cancer cells [J]. The New England journal of medicine,2008,359(4):366-77.
[23]NAGRATH S, SEQUIST L V, MAHESWARAN S, et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology [J]. Nature, 2007,450(7173):1235-9.
[24]HELZER K T, BARNES H E, DAY L, et al. Circulating tumor cells are transcriptionally similar to the primary tumor in a murine prostate model [J]. Cancer research,2009,69(19):7860-6.
[25]VREDENBURGH J J, SILVA O, TYER C, et al. A comparison of immunohistochemistry, two-color immunofluorescence, and flow cytometry with cell sorting for the detection of micrometastatic breast cancer in the bone marrow [J]. Journal of hematotherapy,1996,5(1):57-62.
[26]ALIX-PANABIERES C, VENDRELL J P, SLIJPER M, et al. Full-length cytokeratin-19 is released by human tumor cells:a potential role in metastatic progression of breast cancer [J]. Breast cancer research:BCR,2009,11(3):R39.
[27]SCHWARZENBACH H, ALIX-PANABIERES C, MULLER I, et al. Cell-free tumor DNA in blood plasma as a marker for circulating tumor cells in prostate cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2009,15(3):1032-8.
[28]APOSTOLOPOULOS C, GIAMAS G, STEBBING J. Clinical significance of circulating tumor cells [J]. Biomarkers in medicine,2007,1(3):341-2.
[29]ALIX-PANABIERES C, VENDRELL J P, PELLE O, et al. Detection and characterization of putative metastatic precursor cells in cancer patients [J]. Clinical chemistry,2007,53(3):537-9.
[30]ALIX-PANABIERES C, REBILLARD X, BROUILLET J P, et al. Detection of circulating prostate-specific antigen-secreting cells in prostate cancer patients [J]. Clinical chemistry,2005,51(8):1538-41.
[31]PACHMANN K, CAMARA O, KAVALLARIS A, et al. Monitoring the response of circulating epithelial tumor cells to adjuvant chemotherapy in breast cancer allows detection of patients at risk of early relapse [J]. Journal of clinical oncology:official journal of the American Society of Clinical Oncology,2008, 26(8):1208-15.
[32]PACHMANN K, CLEMENT J H, SCHNEIDER C P, et al. Standardized quantification of circulating peripheral tumor cells from lung and breast cancer [J]. Clinical chemistry and laboratory medicine:CCLM/FESCC,2005,43(6): 617-27.
[33]YAMASHITA J, MATSUO A, KURUSU Y, et al. Preoperative evidence of circulating tumor cells by means of reverse transcriptase-polymerase chain reaction for carcinoembryonic antigen messenger RNA is an independent predictor of survival in non-small cell lung cancer:a prospective study [J]. The Journal of thoracic and cardiovascular surgery,2002,124(2):299-305.
[34]SHER Y P, SHIH J Y, YANG P C, et al. Prognosis of non-small cell lung cancer patients by detecting circulating cancer cells in the peripheral blood with multiple marker genes [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2005,11(1):173-9.
[35]CHEN T F, JIANG G L, FU X L, et al. CK19 mRNA expression measured by reverse-transcription polymerase chain reaction (RT-PCR) in the peripheral blood of patients with non-small cell lung cancer treated by chemo-radiation:an independent prognostic factor [J]. Lung Cancer,2007,56(1):105-14.
[36]MENG S, TRIPATHY D, FRENKEL E P, et al. Circulating tumor cells in patients with breast cancer dormancy [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2004,10(24): 8152-62.
[37]EDERER F, GOLDBLATT S A, NADEL E M. Analysis of the Color Micrograph Study of the Circulating Cancer Cell Cooperative (Cccc) [J]. Acta cytologica, 1965,9(50-7.
[38]LIU L, LIAO G Q, HE P, et al. Detection of circulating cancer cells in lung cancer patients with a panel of marker genes [J]. Biochemical and biophysical research communications,2008,372(4):756-60.
[39]SHEU C C, CHANG M Y, CHANG H C, et al. Combined detection of CEA, CK-19 and c-met mRNAs in peripheral blood:a highly sensitive panel for potential molecular diagnosis of non-small cell lung cancer [J]. Oncology,2006, 70(3):203-11.
[40]YAMAMOTO O, TAKAHASHI H, HIRASAWA M, et al. Surfactant protein gene expressions for detection of lung carcinoma cells in peripheral blood [J]. Respiratory medicine,2005,99(9):1164-74.
[41]ROSS A A, COOPER B W, LAZARUS H M, et al. Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques [J]. Blood,1993, 82(9):2605-10.
[42]MOSTERT B, SLEIJFER S, FOEKENS J A, et al. Circulating tumor cells (CTCs):detection methods and their clinical relevance in breast cancer [J]. Cancer treatment reviews,2009,35(5):463-74.
[43]SHIMOMURA O. Discovery of green fluorescent protein (GFP) (Nobel Lecture) [J]. Angew Chem Int Ed Engl,2009,48(31):5590-602.
[44]CORMACK B P, VALDIVIA R H, FALKOW S. FACS-optimized mutants of the green fluorescent protein (GFP) [J]. Gene,1996,173(1 Spec No):33-8.
[45]ZABAGLO L, ORMEROD M G, PARTON M, et al. Cell filtration-laser scanning cytometry for the characterisation of circulating breast cancer cells [J]. Cytometry Part A:the journal of the International Society for Analytical Cytology,2003,55(2):102-8.
[46]DE GRAAFF M A, CLETON-JANSEN A M, SZUHAI K, et al. Mediator complex subunit 12 exon 2 mutation analysis in different subtypes of smooth muscle tumors confirms genetic heterogeneity [J]. Human pathology,2013,
[47]WANG Z A, MITROFANOVA A, BERGREN S K, et al. Lineage analysis of basal epithelial cells reveals their unexpected plasticity and supports a cell-of-origin model for prostate cancer heterogeneity [J]. Nature cell biology, 2013,15(3):274-83.
[48]MANI S A, GUO W, LIAO M J, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells [J]. Cell,2008,133(4):704-15.
[49]BERX G, RASPE E, CHRISTOFORI G, et al. Pre-EMTing metastasis? Recapitulation of morphogenetic processes in cancer [J]. Clinical & experimental metastasis,2007,24(8):587-97.
[50]CHENG W Y, KANDEL J J, YAMASHIRO D J, et al. A multi-cancer mesenchymal transition gene expression signature is associated with prolonged time to recurrence in glioblastoma [J]. PloS one,2012,7(4):e34705.
[51]STOYIANNI A, GOUSSIA A, PENTHEROUDAKIS G, et al. Immunohistochemical study of the epithelial-mesenchymal transition phenotype in cancer of unknown primary:incidence, correlations and prognostic utility [J]. Anticancer research,2012,32(4):1273-81.
[52]SUGIMOTO H, ITO M, HATANO M, et al. Roles of epithelial-mesenchymal transition in squamous cell carcinoma of the temporal bone [J]. Otology & neurotology:official publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology, 2011,32(3):483-7.
[53]PINZANI P, SALVADORI B, SIMI L, et al. Isolation by size of epithelial tumor cells in peripheral blood of patients with breast cancer:correlation with real-time reverse transcriptase-polymerase chain reaction results and feasibility of molecular analysis by laser microdissection [J]. Human pathology,2006,37(6): 711-8.
[54]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Application of a filtration- and isolation-by-size technique for the detection of circulating tumor cells in cutaneous melanoma [J]. The Journal of investigative dermatology,2010, 130(10):2440-7.
[55]HOFMAN V, ILIE M I, LONG E, et al. Detection of circulating tumor cells as a prognostic factor in patients undergoing radical surgery for non-small-cell lung carcinoma:comparison of the efficacy of the CellSearch Assay and the isolation by size of epithelial tumor cell method [J]. International journal of cancer Journal international du cancer,2011,129(7):1651-60.
[56]HOFMAN V, BONNETAUD C, ILIE M I, et al. Preoperative circulating tumor cell detection using the isolation by size of epithelial tumor cell method for patients with lung cancer is a new prognostic biomarker [J]. Clinical cancer research:an official journal of the American Association for Cancer Research, 2011,17(4):827-35.
[57]HOFMAN V J, ILIE M I, BONNETAUD C, et al. Cytopathologic detection of circulating tumor cells using the isolation by size of epithelial tumor cell method: promises and pitfalls [J]. American journal of clinical pathology,2011,135(1): 146-56.
[58]KIDD J G, ROUS P. Cancers Deriving from the Virus Papillomas of Wild Rabbils under Natural Conditions [J]. The Journal of experimental medicine, 1940,71(4):469-94.
[59]GIRI I, DANOS O, YANIV M. Genomic structure of the cottontail rabbit (Shope) papillomavirus [J]. Proceedings of the National Academy of Sciences of the United States of America,1985,82(6):1580-4.
[60]WOYWODT A, GOLDBERG C, SCHEER J, et al. An improved assay for enumeration of circulating endothelial cells [J]. Annals of hematology,2004, 83(8):491-4.
[61]KREBS M G., HOU J M, SLOANE R, et al. Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and-independent approaches [J]. Journal of thoracic oncology:official publication of the International Association for the Study of Lung Cancer,2012,7(2): 306-15.
[62]WERLING R W, YAZIJI H, BACCHI C E, et al. CDX2, a highly sensitive and specific marker of adenocarcinomas of intestinal origin:an immunohistochemical survey of 476 primary and metastatic carcinomas [J]. The American journal of surgical pathology,2003,27(3):303-10.
[63]BOCKHORN M, JAIN R K, MUNN L L. Active versus passive mechanisms in metastasis:do cancer cells crawl into vessels, or are they pushed? [J]. The lancet oncology,2007,8(5):444-8.
[64]CRISTOFANILLI M, BUDD G T, ELLIS M J, et al. Circulating tumor cells, disease progression, and survival in metastatic breast cancer [J]. The New England journal of medicine,2004,351(8):781-91.
[65]BUDD G T, CRISTOFANILLI M, ELLIS M J, et al. Circulating tumor cells versus imaging--Predicting overall survival in metastatic breast cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(21):6403-9.
[66]PIERGA J Y, BIDARD F C, MATHIOT C, et al. Circulating tumor cell detection predicts early metastatic relapse after neoadjuvant chemotherapy in large operable and locally advanced breast cancer in a phase II randomized trial [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2008,14(21):7004-10.
[67]PESTRIN M, BESSIS, GALARDI F, et al. Correlation of HER2 status between primary tumors and corresponding circulating tumor cells in advanced breast cancer patients [J]. Breast cancer research and treatment,2009,118(3):523-30.
[68]KALLURI R, WEINBERG R A. The basics of epithelial-mesenchymal transition [J]. The Journal of clinical investigation,2009,119(6):1420-8.
[69]THIERY J P. Epithelial-mesenchymal transitions in tumour progression [J]. Nature reviews Cancer,2002,2(6):442-54.
[70]THIERY J P, ACLOQUE H, HUANG R Y, et al. Epithelial-mesenchymal transitions in development and disease [J]. Cell,2009,139(5):871-90.
[71]MEGO M, MANI S A, CRISTOFANILLI M. Molecular mechanisms of metastasis in breast cancer--clinical applications [J]. Nature reviews Clinical oncology,2010,7(12):693-701.
[72]ALLARD W J, MATERA J, MILLER M C, et al. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2004,10(20):6897-904.
[73]FARACE F, MASSARD C, VIMOND N, et al. A direct comparison of CellSearch and ISET for circulating tumour-cell detection in patients with metastatic carcinomas [J]. British journal of cancer,2011,105(6):847-53.
[74]KHOJA L, BACKEN A, SLOANE R, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker [J]. British journal of cancer,2012,106(3):508-16.
[75]LIOTTA L A, KLEINERMAN J, SAIDEL G M. Quantitative relationships of intravascular tumor cells, tumor vessels, and pulmonary metastases following tumor implantation [J]. Cancer research,1974,34(5):997-1004.
[76]WEITZ J, KIENLE P, LACROIX J, et al. Dissemination of tumor cells in patients undergoing surgery for colorectal cancer [J]. Clinical cancer research: an official journal of the American Association for Cancer Research,1998,4(2): 343-8.
[77]ESCHWEGE P, MOUTEREAU S, DROUPY S, et al. Prognostic value of prostate circulating cells detection in prostate cancer patients:a prospective study [J]. British journal of cancer,2009,100(4):608-10.
[78]SCHMIDT T, KOCH M, ANTOLOVIC D, et al. Influence of two different resection techniques (conventional liver resection versus anterior approach) of liver metastases from colorectal cancer on hematogenous tumor cell dissemination-prospective randomized multicenter trial [J]. BMC surgery,2008, 8(6.
[79]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Circulating benign nevus cells detected by ISET technique:warning for melanoma molecular diagnosis [J]. Archives of dermatology,2010,146(10):1120-4.
[1]WORLD HEALTH ORGANIZATION. The global burden of disease:2004 update [M]. Geneva: World Health Organization,2008.
[2]KOHN E C, LIOTTA L A. Molecular insights into cancer invasion:strategies for prevention and intervention [J]. Cancer research,1995,55(9):1856-62.
[3]FIDLER I J. Tumor heterogeneity and the biology of cancer invasion and metastasis [J]. Cancer research,1978,38(9):2651-60.
[4]GUPTA G P, MASS AGUE J. Cancer metastasis: building a framework [J]. Cell, 2006,127(4):679-95.
[5]MOSTERT B, SLEIJFER S, FOEKENS J A, et al. Circulating tumor cells (CTCs): detection methods and their clinical relevance in breast cancer [J]. Cancer treatment reviews,2009,35(5):463-74.
[6]ROSS A A, COOPER B W, LAZARUS H M, et al. Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques [J]. Blood,1993, 82(9):2605-10.
[7]VONA G, SABILE A, LOUHA M, et al. Isolation by size of epithelial tumor cells: a new method for the immunomorphological and molecular characterization of circulatingtumor cells [J]. The American journal of pathology,2000,156(1): 57-63.
[8]MENG S, TRIPATHY D, FRENKEL E P, et al. Circulating tumor cells in patients with breast cancer dormancy [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2004,10(24):8152-62.
[9]ZABAGLO L, ORMEROD M G, PARTON M, et al. Cell filtration-laser scanning cytometry for the characterisation of circulating breast cancer cells [J]. Cytometry Part A:the journal of the International Society for Analytical Cytology,2003,55(2):102-8.
[10]PATERLINI-BRECHOT P, BENALI N L. Circulating tumor cells (CTC) detection: clinical impact and future directions [J]. Cancer letters,2007,253(2): 180-204.
[11]WOYWODT A, GOLDBERG C, SCHEER J, et al. An improved assay for enumeration of circulating endothelial cells [J]. Annals of hematology,2004, 83(8):491-4.
[12]HOFMAN V, BONNETAUD C, ILIE M I, et al. Preoperative circulating tumor cell detection using the isolation by size of epithelial tumor cell method for patients with lung cancer is a new prognostic biomarker [J]. Clinical cancer research:an official journal of the American Association for Cancer Research, 2011,17(4):827-35.
[13]HOFMAN V J, ILIE M I, BONNETAUD C, et al. Cytopathologic detection of circulating tumor cells using the isolation by size of epithelial tumor cell method: promises and pitfalls [J]. American journal of clinical pathology,2011,135(1): 146-56.
[14]KREBS M G, HOU J M, SLOANE R, et al. Analysis of circulating tumor cells in patients with non-small cell lung cancer using epithelial marker-dependent and-independent approaches [J]. Journal of thoracic oncology:official publication of the International Association for the Study of Lung Cancer,2012,7(2): 306-15.
[15]HRISTOZOVA T, KONSCHAK R, BUDACH V, et al. A simple multicolor flow cytometry protocol for detection and molecular characterization of circulating tumor cells in epithelial cancers [J]. Cytometry Part A:the journal of the International Society for Analytical Cytology,2012,
[16]COHEN S J, ALPAUGH R K, GROSS S, et al. Isolation and characterization of circulating tumor cells in patients with metastatic colorectal cancer [J]. Clinical colorectal cancer,2006,6(2):125-32.
[17]STRIJBOS M H, GRATAMA J W, KRAAN J, et al. Circulating endothelial cells in oncology: pitfalls and promises [J]. British journal of cancer,2008,98(11): 1731-5.
[18]SMIRNOV D A, FOULK B W, DOYLE G V, et al. Global gene expression profiling of circulating endothelial cells in patients with metastatic carcinomas [J]. Cancer research,2006,66(6):2918-22.
[19]HOU J M, KREBS M, WARD T, et al. Circulating tumor cells as a window on metastasis biology in lung cancer [J]. The American journal of pathology,2011, 178(3):989-96.
[20]KHOJA L, BACKEN A, SLOANE R, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker [J]. British journal of cancer,2012,106(3):508-16.
[21]VONA G, ESTEPA L, BEROUD C, et al. Impact of cytomorphological detection of circulating tumor cells in patients with liver cancer [J]. Hepatology, 2004,39(3):792-7.
[22]PINZANI P, SALVADORI B, SIMI L, et al. Isolation by size of epithelial tumor cells in peripheral blood of patients with breast cancer: correlation with real-time reverse transcriptase-polymerase chain reaction results and feasibility of molecular analysis by laser microdissection [J]. Human pathology,2006,37(6): 711-8.
[23]PANTEL K, ALIX-PANABIERES C, RIETHDORF S. Cancer micrometastases [J]. Nature reviews Clinical oncology,2009,6(6):339-51.
[24]HAYES D F, CRISTOFANILLI M, BUDD G T, et al. Circulating tumor cells at each follow-up time point during therapy of metastatic breast cancer patients predict progression-free and overall survival [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(14 Pt 1):4218-24.
[25]BIDARD F C, VINCENT-SALOMON A, SIGAL-ZAFRANI B, et al. Prognosis of women with stage IV breast cancer depends on detection of circulating tumor cells rather than disseminated tumor cells [J]. Annals of oncology:official journal of the European Society for Medical Oncology/ESMO,2008,19(3): 496-500.
[26]STOTT S L, LEE R J, NAGRATH S, et al. Isolation and characterization of circulating tumor cells from patients with localized and metastatic prostate cancer [J]. Science translational medicine,2010,2(25):25ra3.
[27]MAHESWARAN S, SEQUIST L V, NAGRATH S, et al. Detection of mutations in EGFR in circulating lung-cancer cells [J]. The New England journal of medicine,2008,359(4):366-77.
[28]NAGRATH S, SEQUIST L V, MAHESWARAN S, et al. Isolation of rare circulating tumour cells in cancer patients by microchip technology [J]. Nature, 2007,450(7173):1235-9.
[29]HELZER K T, BARNES H E, DAY L, et al. Circulating tumor cells are transcriptionally similar to the primary tumor in a murine prostate model [J]. Cancer research,2009,69(19):7860-6.
[30]VREDENBURGH J J, SILVA O, TYER C, et al. A comparison of immunohistochemistry, two-color immunofluorescence, and flow cytometry with cell sorting for the detection of micrometastatic breast cancer in the bone marrow [J]. Journal of hematotherapy,1996,5(1):57-62.
[31]ALIX-PANABIERES C, VENDRELL J P, SLIJPER M, et al. Full-length cytokeratin-19 is released by human tumor cells:a potential role in metastatic progression of breast cancer [J]. Breast cancer research:BCR,2009,11(3):R39.
[32]SCHWARZENBACH H, ALIX-PANABIERES C, MULLER I, et al. Cell-free tumor DNA in blood plasma as a marker for circulating tumor cells in prostate cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2009,15(3):1032-8.
[33]APOSTOLOPOULOS C, GIAMAS G, STEBBING J. Clinical significance of circulating tumor cells [J]. Biomarkers in medicine,2007,1(3):341-2.
[34]ALIX-PANABIERES C, VENDRELL J P, PELLE O, et al. Detection and characterization of putative metastatic precursor cells in cancer patients [J]. Clinical chemistry,2007,53(3):537-9.
[35]ALIX-PANABIERES C, REBILLARD X, BROUILLET J P, et al. Detection of circulating prostate-specific antigen-secreting cells in prostate cancer patients [J]. Clinical chemistry,2005,51(8):1538-41.
[36]PACHMANN K, CAMARA O, KAVALLARIS A, et al. Monitoring the response of circulating epithelial tumor cells to adjuvant chemotherapy in breast cancer allows detection of patients at risk of early relapse [J]. Journal of clinical oncology:official journal of the American Society of Clinical Oncology,2008, 26(8):1208-15.
[37]PACHMANN K, CLEMENT J H, SCHNEIDER C P, et al. Standardized quantification of circulating peripheral tumor cells from lung and breast cancer [J]. Clinical chemistry and laboratory medicine:CCLM/FESCC,2005,43(6): 617-27.
[38]LU J, FAN T, ZHAO Q, et al. Isolation of circulating epithelial and tumor progenitor cells with an invasive phenotype from breast cancer patients [J]. International journal of cancer Journal international du cancer,2010,126(3): 669-83.
[39]PANTEL K, BRAKENHOFF R H, BRANDT B. Detection, clinical relevance and specific biological properties of disseminating tumour cells [J]. Nature reviews Cancer,2008,8(5):329-40.
[40]BENOY I H, ELST H, PHILIPS M, et al. Real-time RT-PCR detection of disseminated tumour cells in bone marrow has superior prognostic significance in comparison with circulating tumour cells in patients with breast cancer [J]. British journal of cancer,2006,94(5):672-80.
[41]SCHMITT M, FOEKENS J A. Circulating tumor cells in blood of primary breast cancer patients assessed by a novel RT-PCR test kit and comparison with status of bone marrow-disseminated tumor cells [J]. Breast cancer research:BCR, 2009,11(5):109.
[42]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Application of a filtration-and isolation-by-size technique for the detection of circulating tumor cells in cutaneous melanoma [J]. The Journal of investigative dermatology,2010, 130(10):2440-7.
[43]HOFMAN V, ILIE M I, LONG E, et al. Detection of circulating tumor cells as a prognostic factor in patients undergoing radical surgery for non-small-cell lung carcinoma:comparison of the efficacy of the CellSearch Assay and the isolation by size of epithelial tumor cell method [J]. International journal of cancer Journal international du cancer,2011,129(7):1651-60.
[44]DE ALBUQUERQUE A, KUBISCH I, BREIER G, et al. Multimarker gene analysis of circulating tumor cells in pancreatic cancer patients:a feasibility study [J]. Oncology,2012,82(1):3-10.
[45]SALOUSTROS E, PERRAKI M, APOSTOLAKI S, et al. Cytokeratin-19 mRNA-positive circulating tumor cells during follow-up of patients with operable breast cancer:prognostic relevance for late relapse [J]. Breast cancer research:BCR,2011,13(3):R60.
[46]DE GIORGI V, PINZANI P, SALVIANTI F, et al. Circulating benign nevus cells detected by ISET technique:warning for melanoma molecular diagnosis [J]. Archives of dermatology,2010,146(10):1120-4.
[47]SCHUSTER R, BECHRAKIS N E, STROUX A, et al. Prognostic relevance of circulating tumor cells in metastatic uveal melanoma [J]. Oncology,2011, 80(1-2):57-62.
[48]PINZANI P, MAZZINI C, SALVIANTI F, et al. Tyrosinase mRNA levels in the blood of uveal melanoma patients:correlation with the number of circulating tumor cells and tumor progression [J]. Melanoma research,2010,20(4):303-10.
[49]PECK K, SHER Y P, SHIH J Y, et al. Detection and quantitation of circulating cancer cells in the peripheral blood of lung cancer patients [J]. Cancer research, 1998,58(13):2761-5.
[50]PANTELEAKOU Z, LEMBESSIS P, SOURLA A, et al. Detection of circulating tumor cells in prostate cancer patients:methodological pitfalls and clinical relevance [J]. Mol Med,2009,15(3-4):101-14.
[51]KUTUN S, CELIK A, CEM KOCKAR M, et al. Expression of CK-19 and CEA mRNA in peripheral blood of gastric cancer patients [J]. Experimental oncology, 2010,32(4):263-8.
[52]LANKIEWICZ S, ROTHER E, ZIMMERMANN S, et al. Tumour-associated transcripts and EGFR deletion variants in colorectal cancer in primary tumour, metastases and circulating tumour cells [J]. Cellular oncology:the official journal of the International Society for Cellular Oncology,2008,30(6):463-71.
[53]BECKER S, BECKER-PERGOLA G, FEHM T, et al. Time is an important factor when processing samples for the detection of disseminated tumor cells in blood/bone marrow by reverse transcription-PCR [J]. Clinical chemistry,2004, 50(4):785-6.
[54]SASTRE J, MAESTRO M L, PUENTE J, et al. Circulating tumor cells in colorectal cancer:correlation with clinical and pathological variables [J]. Annals of oncology:official journal of the European Society for Medical Oncology/ ESMO,2008,19(5):935-8.
[55]DE BONO J S, SCHER H I, MONTGOMERY R B, et al. Circulating tumor cells predict survival benefit from treatment in metastatic castration-resistant prostate cancer [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2008,14(19):6302-9.
[56]RIETHDORF S, FRITSCHE H, MULLER V, et al. Detection of circulating tumor cells in peripheral blood of patients with metastatic breast cancer:a validation study of the CellSearch system [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2007,13(3): 920-8.
[57]FARACE F, MASSARD C, VIMOND N, et al. A direct comparison of CellSearch and ISET for circulating tumour-cell detection in patients with metastatic carcinomas [J]. British journal of cancer,2011,105(6):847-53.
[58]MEDINA A, BROWN E, CARR N, et al. Circulating monocytes have the capacity to be transdifferentiated into keratinocyte-like cells [J]. Wound repair and regeneration:official publication of the Wound Healing Society [and] the European Tissue Repair Society,2009,17(2):268-77.
[59]CHENG W Y, KANDEL J J, YAMASHIRO D J, et al. A multi-cancer mesenchymal transition gene expression signature is associated with prolonged time to recurrence in glioblastoma [J]. PloS one,2012,7(4):e34705.
[60]STOYIANNI A, GOUSSIA A, PENTHEROUDAKIS G, et al. Immunohistochemical study of the epithelial-mesenchymal transition phenotype in cancer of unknown primary:incidence, correlations and prognostic utility [J]. Anticancer research,2012,32(4):1273-81.
[61]SUGIMOTO H, ITO M, HATANO M, et al. Roles of epithelial-mesenchymal transition in squamous cell carcinoma of the temporal bone [J]. Otology & neurotology:official publication of the American Otological Society, American Neurotology Society [and] European Academy of Otology and Neurotology, 2011,32(3):483-7.
[62]BERX G, RASPE E, CHRISTOFORI G, et al. Pre-EMTing metastasis? Recapitulation of morphogenetic processes in cancer [J]. Clinical & experimental metastasis,2007,24(8):587-97.
[63]KALLURI R, WEINBERG R A. The basics of epithelial-mesenchymal transition [J]. The Journal of clinical investigation,2009,119(6):1420-8.
[64]THIERY J P. Epithelial-mesenchymal transitions in tumour progression [J]. Nature reviews Cancer,2002,2(6):442-54.
[65]MANI S A, GUO W, LIAO M J, et al. The epithelial-mesenchymal transition generates cells with properties of stem cells [J]. Cell,2008,133(4):704-15.
[66]POLYAK K, WEINBERG R A. Transitions between epithelial and mesenchymal states:acquisition of malignant and stem cell traits [J]. Nature reviews Cancer,2009,9(4):265-73.
[67]THIERY J P, ACLOQUE H, HUANG R Y, et al. Epithelial-mesenchymal transitions in development and disease [J]. Cell,2009,139(5):871-90.
[68]LECHARPENTIER A, VIELH P, PEREZ-MORENO P, et al. Detection of circulating tumour cells with a hybrid (epithelial/mesenchymal) phenotype in patients with metastatic non-small cell lung cancer [J]. British journal of cancer, 2011,105(9):1338-41.
[69]OZAKI K, YAMAGAMI T, NOMURA K, et al. Prognostic significance of surgical margin, Ki-67 and cyclin D1 protein expression in grade II canine cutaneous mast cell tumor [J]. The Journal of veterinary medical science/the Japanese Society of Veterinary Science,2007,69(11):1117-21.
[70]BALIC M, LIN H, YOUNG L, et al. Most early disseminated cancer cells detected in bone marrow of breast cancer patients have a putative breast cancer stem cell phenotype [J]. Clinical cancer research:an official journal of the American Association for Cancer Research,2006,12(19):5615-21.
[71]ALLAN A L, VANTYGHEM S A, TUCK A B, et al. Tumor dormancy and cancer stem cells:implications for the biology and treatment of breast cancer metastasis [J]. Breast disease,2006,26(87-98.
[72]HOFMAN V, LONG E, ILIE M, et al. Morphological analysis of circulating tumour cells in patients undergoing surgery for non-small cell lung carcinoma using the isolation by size of epithelial tumour cell (ISET) method [J]. Cytopathology:official journal of the British Society for Clinical Cytology, 2012,23(1):30-8.
[73]MAHESWARAN S, HABER D A. Circulating tumor cells:a window into cancer biology and metastasis [J]. Current opinion in genetics & development, 2010,20(1):96-9.
[74]LELIEVRE L, PATERLINI-BRECHOT P, CAMATTE S, et al. Effect of laparoscopy versus laparotomy on circulating tumor cells using isolation by size of epithelial tumor cells [J]. International journal of gynecological cancer official journal of the International Gynecological Cancer Society,2004,14(2): 229-33.
[75]NANAL R, KYLE P, SOOTHILL P W. A classification of pregnancy losses after invasive prenatal diagnostic procedures:an approach to allow comparison of units with a different case mix [J]. Prenatal diagnosis,2003,23(6):488-92.
[76]BIANCHI D W. Fetal cells in the maternal circulation:feasibility for prenatal diagnosis [J]. British journal of haematology,1999,105(3):574-83.
[77]NICE. Antenatal care-routine care for the healthy pregnant woman-clinical guideline [M]. London:RCOG Press (NICE Clinical Guidelines, No.62.).2008 Mar.
[78]FINNING K M, MARTIN P G, SOOTHILL P W, et al. Prediction of fetal D status from maternal plasma:introduction of a new noninvasive fetal RHD genotyping service [J]. Transfusion,2002,42(8):1079-85.
[79]VONA G, BEROUD C, BENACHI A, et al. Enrichment, immunomorphological, and genetic characterization of fetal cells circulating in maternal blood [J]. The American journal of pathology,2002,160(1):51-8.
[80]BEROUD C, KARLIOVA M, BONNEFONT J P, et al. Prenatal diagnosis of spinal muscular atrophy by genetic analysis of circulating fetal cells [J]. Lancet, 2003,361(9362):1013-4.
[81]SAKER A, BENACHI A, BONNEFONT J P, et al. Genetic characterisation of circulating fetal cells allows non-invasive prenatal diagnosis of cystic fibrosis [J]. Prenatal diagnosis,2006,26(10):906-16.
[82]FUNAKI S, SAWABATA N, NAKAGIRI T, et al. Novel approach for detection of isolated tumor cells in pulmonary vein using negative selection method: morphological classification and clinical implications [J]. European journal of cardio-thoracic surgery:official journal of the European Association for Cardio-thoracic Surgery,2011,40(2):322-7.
[83]MEHES G, WITT A, KUBISTA E, et al. Circulating breast cancer cells are frequently apoptotic [J]. The American journal of pathology,2001,159(1):17-20.
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