液体活检专业知识宣讲培训课件.ppt

单击此处编辑母版标题样式,单击此处编辑母版文本样式,第二级,第三级,第四级,第五级,*,本文档所提供的信息仅供参考之用，不能作为科学依据，请勿模仿；如有不当之处，请联系网站或本人删除。,*,*,本文档所提供的信息仅供参考之用，不能作为科学依据，请勿模仿；如有不当之处，请联系网站或本人删除。,内容摘要,一,.,液体活检,1,）,cfDNA&ctDNA&CTC,概念和最新研究进展,二,.CTC/CTSC,研究方法和进展,2）液体活检应用和意义,三,.,我们的研究,液态活检,来源于肿瘤组织，存在于血液中：,循环肿瘤细胞（,CTC,）,循环肿瘤,DNA,（,ctDNA,）,肿瘤外泌体,exosome,提示肿瘤发展进程及抗药性等信息，指导个体化精准治疗,Diaz LA Jr,Bardelli A.,Liquid Biopsies:Genotyping Circulating Tumor DNA,.J Clin Oncol.2014;32(6):579586,cfDNA,What,:,循环游离,DNA,，机体细胞释放入外周血循环后发生部分降解的内源性,DNA,。高度片段化，约,170bp,。,Source:,正常细胞、异常细胞（如肿瘤细胞），病毒,DNA,等,State:,半衰期短，动态表达量，核小体以单个、双联或三联的形式进入血液，并逐步分解。,cfDNA,（,cell-free DNA,）,ctDNA,What,:,循环肿瘤,DNA,，是指肿瘤细胞,DNA,经脱落或细胞凋亡后释放进入循环系统。一种特征性的,肿瘤生物标记物,，可以被定性、定量和追踪。,Source,:,来源于肿瘤细胞的,cfDNA,，属于,cfDNA,的一种类型。不同患者的水平差异显著，受肿瘤负荷，分期等因素的影响,ctDNA,（,circulating tumor DNA,）,cfDNA&ctDNA,Ellen Heitzer,Peter Ulz,and Jochen B.Geig.,Circulating Tumor DNA as a Liquid Biopsy for Cancer.Clinical Chemistry.2015,112123,1,：,CAPP-Seq,（,cancer personalized profiling by deep sequencing,）癌症个体化深度测序分析方法,:,利用定制化的,NimbleGen SeqCap EZ Choice Library,作为”筛选器“对样本靶向捕获后再深度测序，灵敏度高，特异性强且经济可行。,2,：,利用数字错误抑制（,IDES,）的方法和,CAPP-seq,（,ctDNA,测序方法），能够检测到频率低至,0.004,的突变等位基因,Newman AM,Lovejoy A F ,Klass DM,et al.Integrated digital error suppression for improved detection of circulating tumor DNA.Nature Biotechnology(2016)doi:10.1038/nbt.3520,ctDNA-,高通量测序,cfDNA,片段谱系分析,Deep sequencing of circulating cell-free,DNA yields a dense,genome-wide map of,nucleosome occupancy that enables identication of its,cell-types of origin,potentially enabling the noninvasive,monitoring,of a broader set of,clinical conditions,Matthew W.Snyder,Martin Kircher,Andrew J.Hillet al.Cell-free DNA Comprises an In Vivo Nucleosome Footprint that Informs Its Tissues-Of-Origin.Cell.2016 Jan 14;164(1-2):57-68.,CTC,What,：,循环肿瘤细胞，是指从实体瘤中脱离出来并进入外周血液循环的肿瘤细胞。,Source:,原发肿瘤,和,转移灶,都会释放,CTC,。可检测到的,CTC,是肿瘤细胞在循环系统微环境下生存竞争的结果,。,CTC,（,circulating tumor cells,）,液体活检应用价值,（监测肿瘤特异性突变发现肿瘤复发）,Ellen Heitzer,Peter Ulz,and Jochen B.Geig.Circulating Tumor DNA as a Liquid Biopsy for Cancer.Clinical Chemistry.2015,112123,液体活检应用价值,（,分析耐药株的遗传变异）,Ellen Heitzer,Peter Ulz,and Jochen B.Geig.,Circulating Tumor DNA as a Liquid Biopsy for Cancer.Clinical Chemistry.2015,112123,液体活检优势及前景,液态活检无侵入性、可频繁多次检测及快速反应能力均体现出发展潜力巨大应用价值。将削减无效肿瘤医疗开支、降低医疗成本。,适应症广泛，如,乳腺癌、结直肠癌、肺癌、胃癌、食管癌,等常见肿瘤均可用液态活检技术进行诊断与监测。,2015,年美国发布的肿瘤治疗计划的四大举措之一：“,液态活检,”血浆开发新方法来评估治疗反应以及抵抗可能的耐药性。,根据中国国家癌症中心发布的数据，我国,5,年内诊断为癌症且仍存活的病例数约为,749,万,。也是液态活检巨大的市场。,CTC,研究概况,1,）,CTC,与转移的关系,2）CTC异质性与肿瘤治疗,3）CTC应用和意义,4）CTC 研究方法,CTCs,(circulating tumor cells,CTCs),Ashworth,早在,1896,年就提出了循环肿瘤细胞的概念。侵入循环系统的肿瘤细胞，大部分由于机体的免疫识别、机械杀伤及自身凋亡在短期内死亡，只有极少数存活下来，在适于自身存活和增殖的器官或组织形成转移灶。,CTC,与肿瘤转移,Simon A Joosse,Tobias M Gorges,and Klaus Pantel.Biology,detection,and clinical implications of circulating tumor cells.EMBO Mol Med.2015;7(1):111.,Brouwer A,De Laere B,Peeters D,et al.,consequences of heterogeneity in the circulating tumor cell compartment.,Oncotarget.2016 Mar 9.doi:10.18632/oncotarget.8015.,CTC,异质性,CTC,应用和意义,CTCs,肿瘤转移预测,肿瘤治疗,CTCs,数量改变反映了肿瘤细胞对各种治疗方案的敏感性和有效性，为个体化治疗提供依据,。,监测早期转移,乳腺癌、前列腺癌,结肠癌、肺癌、胃癌、肝癌、胰腺癌多种肿瘤,监测,CTCs,基因表达突变，找到药物敏感的靶点，指导个体化治疗,CTC,优化治疗方案,2014,年,GoldKorn,，接受,多西他赛联合阿曲生坦治疗,的转移性去势抵抗性前列腺癌患者，基线循环肿瘤细胞,CTC,数量可预测预后，,3,周时,CTC,升高提示总生存较差，有望作为,重新指导,和,优化治疗,的较好指标,。,Goldkorn A,，,Ely B,，,Quinn DI,，,et al,Circulating Tumo cell counts are Prognostic of overall survival in SWOG S0421,：,a Phase III Trial of docetaxel with or without atrasentan For Metastatic castration resistant prostate cancer.J Clin Oncol2014 32(11),：,1136-1142,2012,年,Bian,乳癌患者在接受化疗,（,白蛋白结合型紫杉醇,+,卡培他滨,+,曲妥珠单抗）前后,CTC,变化。,治疗前,：,CTC,3,，,HER-2+,；,1,疗程：,CTC 0,。,6,疗程：,病理学,CR,。,证实,CTC,值在乳腺癌,治疗早期,发生显著变化，能够准确,预示疗效，能够科学指导选择治疗方案,。,Bian L,，,Wang T,，,Liu Y,，,et al,Evaluation of treatment response for breast cancer,：,are we Entering the era of”biological Complete remission”?,Chin J Cancer Res,2012,，,24,：,403-407,CTCs,的研究方法,Ramdane Harouaka,Zhigang Kang,Si-Yang Zheng,Liang Cao.Circulating tumor cells:Advances in isolation and analysis,and challenges for clinical applications.Pharmacology 141:209221.,研究方法,（,1,）,-Cell Search,CellSearch,系统是,唯一,经,FDA,批准进入临床应用的,CTCs,检测方案。,原理：,EpCAM,抗体标识的微磁珠从全血中捕获细胞，用免疫荧光识,EpCAM+CK+CD45,进行磁性分离,。,CTC,EpCAM,CK,CD45,DAPI,Cell Search,数据解读,:,1.7.5ML,外周血,判断,阈值：,转移性,乳腺癌,CTCs,5,细胞,前列腺癌,CTCs,5,细胞,结肠癌,：,CTCs,3,细胞,表明,预后不良,3.,目前,FDA,只,批准用于以上,三种,肿瘤,CTCs,的研究方法（,2,）,-,基因芯片,CTCs,的研究方法（,3,）,-,测序,Xiaohui Nia,Minglei Zhuo,Zhe Sua,，,et al.Reproducible copy number variation patterns among single circulating tumor cells of lung cancer patients.PNA,2013,52(110)21083-21088.,Xiaohui Nia,Minglei Zhuo,Zhe Sua,，,et al.Reproducible copy number variation patterns among single circulating tumor cells of lung cancer patients.PNA,2013,52(110)21083-21088.,Ramdane Harouaka,Zhigang Kang,Si-Yang Zheng,Liang Cao.Circulating tumor cells:Advances in isolation and analysis,and challenges for clinical applications.Pharmacology 141:209221.,CTCs,的分离方法（,1,）,CTCs,的分离方法（,2,）,-,体外培养,Min Yu,Aditya Bardia,Nicola Aceto,et al.Ex vivo culture of circulating breast tumor cells for,individualized testing of drug susceptibility.,Science.,2014;345(6193):216220.,Papanicolaou staining for uncultured primary,Representative images of nonadherent CTC,immunofluorescent staining,：,CK,：,red,；,Ki67,：,yellow,；,CD45,：,green,；,DAPI,：,blue.,CTSC,研究概况,1,）,CTSC,定义,2）,CSC,CTC,CTSC,三者关系,3,）CT,S,C研究进展,4,）CT,S,C 研究难点和未来方向,CTSC,背景介绍,循环肿瘤干细胞（,circulating tumor stem cells,CTSCs,）,：,CTCs,中具有,干细胞特性,，即自我更新及分化潜能的一小群细胞,。,作为标志物,CTSC,具有更重要的临床价值。,EMT,可使,CTCs,获得干细胞特性，而具有,药物耐受性,和,高侵袭性,。,CTSC,已在包括肝癌，乳腺癌，胃癌，结直肠癌等多种肿瘤中被证实。,CSC,CTC,CTSC,（,肿瘤转移等级结构图,）,Yang MH,Imrali A,Heeschen C.Circulating cancer stem cells:the importance to select.Chin J Cancer Res.2015 Oct;27(5):437-49.,Markers for CTSC,Yang MH,Imrali A,Heeschen C.Circulating cancer stem cells:the importance to select.Chin J Cancer Res.2015 Oct;27(5):437-49.,乳腺癌,CTSC,研究进展,学者们采用不同的表面标志物组合来定义,CTSC,：,Epcam/CK/CD44,CD24,；,Epcam/CK/ALDH1,CD24,Epcam/CK/Vimentin/Fibernectin,；,Epcam/CK/TWISTI/Akt2PDKa,Epcam/Muc1/HER2,KasimirBauer S,，,Hoffmann 0,，,Wallwiener D,，,et a1,Expression Of stem cell and epithelialmesenchymal transition markers in Primary breast cancer Patients With circulating tumor cells J,Breast Cancer Res,，,2012,，,14(1),：,R15,Baccelli I,，,Schneeweiss A,，,Riethdorf S,，,et a1,Identification of a population of blood circulating tumor cells from breast cancer patients that initiates metastasis in a xenografi assayJ,Nat Biotechnol,，,2013,，,31(6),：,539,544,乳腺癌,CTSC,研究进展,Yu M,Bardia A,，,Wittner BS,et a1,Circulating breast tumor cells Exhibit dynamic changes In Epithelial and mesenehymal eompostionJ,Science,，,2013,，,339(6119),：,580-584,EPCAM low MET high CD47 high CD44 high,2013,年上海复旦大学附属肝癌研究所樊嘉及周俭研究组通过研究,123,例肝癌患者,CTCs,相关肿瘤干细胞免表标记物，包括,CD133,、,ABCG2,、,CD90,、,-catenin,、,E-cadherin,以及,Vimentin,。,2013,年,Liu,等鉴于,ICAM-1,可能是肝癌干细胞标志物，也采用流式细胞术检测了肝癌血样，发现部分病例可检出,CD45-ICAM-1+,细胞，这样的细胞具有较强的体外成球和体内致瘤能力。,Y.F.,et al.,Circulating stem cell-like epithelial cell adhesion molecule-positive tumor cells indicate poor prognosis of hepatocellular carcinoma after curative resection.Hepatology,2013.57(4):p.1458-68,。,Liu S,Li N,Yu X,et al.Expression of intercellular adhesionmolecule 1 by hepatocellular carcinoma stem cells and circulating tumor cellsJ.Gastroenterology,2013,144(5):1031-1041,肝癌,CTSCs,研究进展,我们的研究,1)CSC and miRNA,2)CTC and CTSC,项目研究基础,前期基金支持,1.,2014,年福建省卫教联合项目：,结直肠癌循环肿瘤细胞和肿瘤干细胞的分子机制研究（,WKJ-FJ-14,）,2.,2012,年福建省科技厅重点项目,:,循环肿瘤细胞与肿瘤干细胞在恶性肿瘤个体化治疗中的应用研究,（,2012Y0017,）；,3.,2012,年福建省卫生厅青年课题,:,结直肠癌循环肿瘤细胞及其上皮间质变标 志物的定量检测,（,2012-2-13,）；,4.,2012,年福建省自然科学基金项目,:miR-148,家族,-,靶蛋白网络在肝癌干细胞中调控机制研究,（,2012J05138,）；,5.,2011,年福建省医学创新课题,:,乳腺癌循环肿瘤细胞（,CTC,）及其单细胞多基因检测的临床意义,（,2011-CX-16,）；,6.,2011,年福建省自然科学基金项目,:,循环肿瘤细胞及其耐药基因表达水平在晚期乳腺癌化疗敏感性中的应用研究,（,2011J01141,）,基金支持,8.,2009,年,福建省自然科学基金项目,:,靶向乳腺癌干细胞的树,突状瘤苗抗肿瘤的研究,(2009J01118),9.,2008,年福建省自然科学基金项目,:,miRNA,靶向肝癌,SP,、,NSP,细胞基因差异表达的研究,（,2008J0077,）,7.2016,年福建省自然科学基金项目：,miR-194,调控肝癌干细,胞的靶基因和,LncRNA,鉴定,（,2016J01436,）,MoFlo,超高速流式细胞分选系统,稀有细胞检测的灵敏度高,分选速度快,研究基础设备,CSC-Side population(SP)assay,It is generally accepted that SP cells possess stem cell characteristics.,Critical:,Optimize Hoechst 33342 staining procedure to achieve a consistent and tumorigenic SP subtype as SP approach is based on the functional property of CSCs to exclude Hoechst-33342-dye via ABC transporters.,SP,NSP,-MoFlo analysis and sorting,-Sphere formation assay,Floating spheres were formed by SP cells,while none by NSP cells,Hep-3B PLC/PRF/5 Huh-7,实验数据,-Tumors formed in NOD/SCID mice,PLC/PRF/5 Huh-7 Hep-3B,Week 12;,110,3,cells;,SP:blue arrow;,NSP:red arrow.,miRNA profiling result by RT-qPCR,miRNA ID,SP/NSP Fold changes,P0.05,miR-9,-3,miR-9,*,-2.41,miR-23b,-2.12,miR-27b,-2.35,miR-96,-2.8,miR-151-3p,-2.65,miR-151-5p,-2.57,miR-183*,-2.23,miR-183,-3.16,miR-194,-2.03,miR-146a,-3.44,miR-148b,-3.14,miR-340-3p,-2.85,miR-421,-2.32,miRNA ID,SP/NSP Fold changes,P0.05,miR-135b*,-3.55,miR-489,-3.28,miR-15a,-2.79,miR-21,-2.67,miR-22,-2.5,miR-365,-2.47,miR-450a-5p,-2.46,miR-7a,-2.44,miR-26b,-2.28,miR-125a-5p,-2.27,miR-25,-2.22,miR-126-3p,-2.15,miR-182,-2.14,A total of,370 miRNA,species were analyzed in PLC/PRF/5 SP and NSP cells,27 miRNAs,were identified as differentially expressed,all of which were down-regulated in SP.,实验数据,-miR-148b,在,SP,中低表达,实验数据,-miR-148b,改变,SP,比例,实验数据,-miR-148b,改变增殖与耐药,实验数据,-miR-148b,改变迁移侵袭与血管生成,实验数据,-miR-148b,过表达抑制肿瘤生成,实验数据,-miR-148b,沉默促进肿瘤生成,研究方案,CTC/CTSC,研究方案,分选流程,已完成,200,例鼻咽癌患者,CTC,检测，,100,例乳腺癌患者,CTC,检测,，,100,例肺癌患者,CTC,检测，,150,例直肠癌,患者,CTC/CTSC,检测，具备,完善的,CTC,检测技术流程,。,在研项目：,150,例初诊,未治肠癌患者，跟踪其全病程临床资料并抽取各阶段血样,，分析,患者,各个时期,CTCs/CTSCs,数值，以,PSF,为观察点，评估,CTCs/CTSCs,数值与,临床分期、预后、治疗方案,及,疗效,等情况的相关性，尤其是,手术配合细胞治疗,的疗效,观察，开发,CTC/CTSC,在,疗效评估中的应用。,研究数据,入组初诊未治肠癌患者,150,例,跟踪患者全程临床资料并抽取各阶段血样,（,TIL,组：,根治术前、术后第一次细胞治疗后、细胞治疗,2,个周期后、细胞治疗,3,个周期后,对照组：,根治术前、术后,7,天、术后,1,月，术后,2,月,每月一次,）,利用,Moflo,对,CTCs/CTSCs,计数,患者各个时期,CTCs/CTSCs,数值与,临床分期、预后、治疗方案及疗效,等情况相关性,以,PSF,为观察点，评估手术配合,细胞治疗,的疗效观察,难点和关键技术,CTC/CTSC,研究难点,稀有性,：仅占,PBMC,的,1/10,6,活性,：难以保持细胞活性进行后续研究,单细胞性,：细胞稀少难以进行后续研究,分选精度可达到,1/10,6,可保持分选后细胞的活性,进行单细胞水平,RNA,提取和预扩增，保证下游基因表达谱分析的质量,MoFlo,超高速流式细胞分选系统,REPLI-g WTA Single,Cell Kit,本项目关键技术,流式分析,CTC,敏感性专一性检测,研究数据,研究数据,CTCs,的画门模式,研究数据,CTSCs,的画门模式,CTCs,的检测,研究数据,研究数据,CTSCs,的检测,首诊初治结直肠癌患者,CTC/CTSC,检测分析结果,Patient Characteristics,Number(%),CTC,、,CTSC(CD44+),p-value,60,25(50.0),13,12,Sex,0.205,female,20(40.0),10,10,man,30(60.0),13,17,Tumor size(cm),0.189,5,26(52.0),13,13,5,24(48.0),10,14,TNM stage,0.072,I,5(10.0),4,1,II,11(22.0),9,2,III,29(58.0),21,8,IV,5(10.0),1,4,T stage,0.118,T1,2(4.0),1,1,T2,5(10.0),5,0,T3,4(8.0),3,1,T4,39(78.0),26,13,RLNM,0.032,N0,17(34.0),14,3,N1,21(42.0),10,11,N2,12(24.0),10,2,M(metastasis),0.024,M0,45(90.0),34,11,M1,5(10.0),1,4,研究数据,结直肠癌患者转移后化疗期间,CTC/CTSC,变化跟踪,研究数据,CTC,分选,分选白细胞对照：,DAPI+/CD45-/Epcam-,分选,CTC,细胞：,DAPI+/CD45-/Epcam+,实验方案,研究数据,-,表达谱芯片聚类图,肿瘤组织 正常组织,CTC,细胞 白细胞对照,实验方法,CNV,区域中,CN,与基因表达的相关分析图,表达谱芯片 比较基因组杂交,近年发表文章,Qinying Liu,Yangmei Xu,Shenghong,et al,.miRNA-148b,suppresses,hepatic cancer stem,cell by targeting neuropilin1.Bioscience Reports,2015;35(4).,Xu,YM,Xie YQ,Wang XR,et al.Identification,of cancer stem cells,from hepetocellular carcinoma cell lines and their related,microRNA,s.,Oncol Rep.,2013.30:2056-62,.,龚福生,，黄伟炜，刘健,等,.,乳腺癌患者循环肿瘤细胞检测的临床意义,.,中华,肿瘤防治杂志,2015,22(13):1009-1013,.,Identification,of perioperative,circulating tumor cells,and potential biomarkers of,circulating tumor stem cells,in colorectal cancer patients by flow cytometry.,待,发表,袁婕,郑秋红 徐鹭英,，等,.,流式细胞术,检测鼻咽癌患者外周血循环肿瘤细胞及临床相关性分析,第八,届中国肿瘤学术大会暨第十三届海峡两岸肿瘤学术会议,论文，,2014.,6.,Zheng,QH,et al.,Floating,cells with,stem cell,properties in gastric cell line SGC-7901.Tumori.2011.,97:393-399.,7.Zheng,QH,XuYM,Wei ZQ,et al.Isolation and purification,of cancer stem-like cells,from cell,line,PLC/PRF-5.,Chinese,Tumor Biological Therapy Journal,2011,18(2):126-132.,8.Lai,L,Cui C,Jin J,et al.,Mouse embryonic stem cel,l-derived thymic epithelial cell,progenitors,enhance T-cell,reconstitution,after allogeneic bone marrow transplantation.Blood.2011.118(12):3410-3418.,近年发表文章,Chen,G,Wang L,Lu J,et al.,Optical Diagnosis for Lung Cancer Using Multiphoton Imaging.,Scanning.,2013.doi:10.1002/sca.21076,.,Yan J,Tan C,Gu F,et al.,Sorafenib delays recurrence and metastasis after liver transplantation in a rat model of hepatocellular carcinoma with high expression of pERK.,Liver Transpl.,2013.doi:10.1002/lt.23619,.,Chen L,Zhang WW,Zheng QH,et al.,Aculeatusquinones A-D,novel metabolites from the marine-derived fungus Aspergillusaculeatus.,Heterocycles.,2013.Published online,.,Zheng QF,Wang JJ,Ying MG,et al.,Omentoplasty in preventing anastomotic leakage of oesophagogastrostomy following radical oesophagectomy with three-field lymphadenectomy.,Eur J Cardiothorac Surg.,2013.43(2):274-8,.,Chen Y,Zheng X,Chen G,et al.,Immunoassay for LMP1 in nasopharyngeal tissue based on surface-enhanced Raman scattering.,Int J Nanomedicine.,2012.7:73-82,.,Yan J,Zhuo S,Chen G,et al.,Preclinical study of using multiphoton microscopy to diagnose liver cancer and differentiate benign and malignant liver lesions.,J Biomed Opt.,2012.17(2):,026004,.,7.Zhuo,S,Yan J,Chen G,et al.,Label-free imaging of basement membranes differentiates normal,precancerous,and cancerous colonic tissues by second-harmonic generation microscopy.,PLoS One.,2012.7(6):e38655.,8.Ying,M,Zhuo S,Chen G,et al.,Real-time noninvasive optical diagnosis for colorectal cancer using multiphoton microscopy.,Scanning.,2012.34(3):181-5,.,未来发展方向,1.,高新技术已,逐渐渗入,CTCs/CTSCs,的,分离与分析,，新研发的技术设备功能更强，性能与检测效果逐步提高。,2.,单细胞全基因测序技术,及,单细胞基因组学,可解决血液,CTCs/CTSCs,数量稀少的难题，能检测更多药效相关基因及分析耐药机制，为,CTCs/CTSCs,的,分子分型及靶向药物治疗,提供更准确的信息。,结语,谢谢！,液体活检，肿瘤精准治疗的宠儿！,CSCs and miRNAs,Takahashi R.,et al.,Front.Genet.2014,2013,年,Campos,等,对,33,例（,0,期,）非转移结直肠癌患者和,9,名健康人，通过,进行,CK,表达、,17,号染色体及,HER2,基因水平分析，,同时,分析,CTC,的表型和基因型,，结果表明：,原发灶无,HER2,蛋白,表达，,却有,63.33%,的,CTC,出现,HER2,蛋白,过,表达。,全面,CTC,分析可以为肿瘤个体化治疗提供更多疾病信息。,Campos M,Luque R,Jimnez J,et al.Simultaneous phenotypic andgenetic characterization of single circulating tumor cells from colon cancer patientsJ.Histol Histopathol,2013,28(11):1439-1450.,CTC,与肿瘤个体化治疗,研究难点,CTC/CTSC,研究难点,稀有性,：仅占,PBMC,的,1/10,6,或者更少,活性,：难以保持细胞活性进行后续研究,单细胞性,：细胞稀少难以进行后续研究,研究数据,-,表达谱芯片散点图,正常组织,肿瘤组织,CTC,细胞,白细胞对照,乳腺癌,CTSC,研究进展,2013,年,Yu,，,侵袭性强的乳腺癌类型，以,间质性表型,CTCs,为主,；而病情恶化,的患者的间质型,CTCs,计数明显增多,。,在,CTCs,动态监测中,，初始对相应靶向药物疗效较,好，,CTCs,数量明显降低并以,上皮型,为主；经,7,个月治疗后出现了耐药性及病情进展,，,间质,CTCs,升高,。,患者,CTCs,、,EMT,干细胞特性与药物疗效或耐药情况密切相关,。,Yu M,Bardia A,，,Wittner BS,et a1,Circulating breast tumor cells Exhibit dynamic changes In Epithelial and mesenehymal eompostionJ,Science,，,2013,，,339(6119),：,580-584,