CN101405399A - 抑制乳腺癌干细胞的生长和转移 - Google Patents
抑制乳腺癌干细胞的生长和转移 Download PDFInfo
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Abstract
披露了抑制表达高分子量－黑色素瘤相关抗原(HMW－MAA)的乳腺癌干细胞生长的方法。该方法包括给予个体一种组合物，该组合物包含有效抑制乳腺癌细胞生长量的与HMW－MAA反应的抗体或这种抗体的片段。还提供了抑制乳腺癌转移的方法或鉴定HMW－MAA+乳腺癌干细胞的方法。
Description
本申请要求2006年3月16日提交的美国临时申请序列号60/783,091的优先权，其内容通过引用纳入本文。
发明领域
本发明一般涉及癌症领域，尤其涉及抑制乳腺癌干细胞的生长。
发明背景
上皮来源的癌症主要造成无法治愈的转移性疾病的癌症相关死亡。癌症干细胞假说(Reya等，(2001)Nature 414：105)认为某些肿瘤的发生和持续是由于组织干细胞的突变，这种突变导致不受控的无限增殖，在这种状态时所述干细胞称为癌症干细胞(CSC)。长久以来都认为只有很少一部分肿瘤细胞能够无限生长(约1/1000到1/5000的肺肿瘤细胞以及1/1,000,000的白血病细胞(Reya，(2001)；Dick，J.E.(2003)Proc Natl Acad Sci 100：3547；Marx，J.(2003)Science301：1308)。现在有了非常好的证据证明，包括乳腺癌(Gudjonsson等，(2002)Genes Dev 16：693；Al-Hajj等，Proc Natl Acad Sci 100：3983；Dontu，G.等，(2004)Breast Cancer Res 6：R605；Ponti，D.，(2005)Cancer Res 65：5506)、结肠癌(Kim等，(2005)Cell 121：823)、卵巢癌(Bapat等，(2005)Cancer Res 65：3025)、肺癌(Kim等，(2005)Cell 121：823)、前列腺癌(Schalken，(2003)Urology62：11)、白血病(Dick，J.E.(2003))、胶质瘤(Kondo等，(2004)Proc Natl AcadSci 101：781；Singh，S.K.等，(2004)Nature 432：396)、视网膜母细胞瘤(Reedijk，M.，S.等，(2005)Cancer Res 65：8530)和肝细胞癌(Rosner，A.，K.等，(2002)Am J Pathol 161：1087)在内的许多癌症是由癌症干细胞增殖造成的。还已经从确定的肿瘤细胞系分离出癌症干细胞(Gudjonsson等，(2002)Genes Dev 16：693；Ponti，D.，(2005)Cancer Res 65：5506；Kondo等，(2004)Proc Natl AcadSci 101：781)，它们保留了与原始分离它们的肿瘤相同的表型。若干癌症类型的证据显示，在正常干细胞功能中显著的途径，尤其是Wht、Notch、Ssh(sonichedgehog)、XIAP(X连锁凋亡抑制蛋白)在癌症干细胞(CSC)中变得“失调”(Reya等，(2001)Nature 414：105；Dontu，G.等，(2004)Breast Cancer Res 6：R605；Rosner，A.，K.等，(2002)Am J Pathol 161：1087；Reya，T.等，(2005)Nature434：843；Li，Y.，B.等，Proc Natl Acad Sci 100：15853；Yang，L.，Z.等，(2003)Cancer Res 63：6815；Liu，S.等，(2005)Breast Cancer Res 7：86；Mikaelian，L等，(2004)Breast Cancer Res 6：R668)。
筛查乳房造影术是鉴定女性乳腺癌非常有效的方法，预计2005年将在美国鉴定出超过211,000例新的侵袭性乳腺癌病例和大约58,000例新的原位乳腺癌病例(Society，A.C.Breast cancer Facts and Figures American Cancer Society2005)。乳腺癌是女性的头号癌症死因(Sasco，A.J.(2003)Horm Res 60增刊3：50)，每年有超过40,000人死于局部和远端转移造成的复发(Society，A.C.Breastcancer Facts and Figures American Cancer Society 2005)。乳腺癌复发与存在全身性微小转移有关。治疗资源非常有限，因为仅能对30％的乳腺癌患者(HER2阳性)联用赫赛汀与放疗和化疗来降低复发率(Bapat，S.A.等，(2005)Cancer Res65：3025)。
即便在手术、化疗、放疗、靶向小分子和抗体疗法之后-所有这些方法都能使肿瘤缩小，但不能消除无限增殖的肿瘤细胞-复发和转移率高都强调需要鉴定特异性针对并杀死癌症干细胞的新治疗策略，以便根治复发性和转移性疾病。因此，目前需要了解仅在癌症干细胞中发生的致瘤性改变可使我们理解CSC肿瘤如何形成、如何增殖、如何逃避标准治疗，还需要开发靶向癌症干细胞的疗法。
发明概述
本发明提供了抑制乳腺癌干细胞生长的方法。所述乳腺癌干细胞表达高分子量-黑色素瘤相关抗原(HMW-MAA)。该方法包括给予个体一种组合物，所述组合物包含有效抑制乳腺癌干细胞生长用量的与HMW-MAA反应的抗体。
在另一实施方式中，提供了一种抑制乳腺癌转移的方法，其中的乳腺癌包含HMW-MAA+乳腺癌干细胞。该方法包括给予个体一种组合物，所述组合物包含一定量的有效抑制乳腺癌转移的与HMW-MAA反应的抗体。
在另一实施方式中，提供了检测HMW-MAA+乳腺癌干细胞的方法。该方法包括将抗体组合物给予个体或使之与获自该个体的生物样品接触，其中所述抗体组合物包含针对HMW-MAA的抗体和至少一种针对乳腺癌干细胞标记物的抗体。同时检测到HMW-MAA抗体和至少一种乳腺癌干细胞标记物的结合可确定存在HMA-MAA+乳腺癌干细胞。
在具体实施方式中，用于实施本发明的抗体可以是指定为225.28的单克隆抗体和/或指定为763.74的单克隆抗体。
附图简述
图1A和1B为通过荧光激活细胞分选(FACS)获得的乳腺癌干细胞某亚群的HMW-MAA表达数据的图示。
图2是MDA-MB-435细胞表达的HMW-MAA的Western印迹分析的照片。
图3A和3B是用乳腺癌干细胞标记物抗体染色的MDA-MB-435s细胞的FACS分离所得数据的图示。
图4是SCID小鼠中HMW-MAA-特异性mAb 763.74和225.28抑制人乳腺癌细胞MDA-MB-435s肺转移所得结果的图解表示。
图5是用mAb 225.28抑制人乳腺癌干细胞术后肺转移所得结果的图解表示。
发明详述
本发明涉及发现乳腺癌干细胞上存在HMW-MAA。本发明提供抑制包含HMW-MAA+乳腺癌干细胞的乳腺癌生长的方法。所述方法包括给予个体一种组合物，所述组合物包含有效抑制乳腺癌干细胞生长用量的与HMW-MAA反应的抗体。
还提供抑制个体乳腺癌转移的方法，其中所述乳腺癌包含HMW-MAA+乳腺癌干细胞。所述方法包括给予该个体一定量的有效抑制转移的与HMW-MAA反应的抗体。
在另一实施方式中，提供检测HMW-MAA+乳腺癌干细胞的方法，该方法通过将抗体组合物给予个体或使之与获自该个体的生物样品接触。所述抗体组合物包含针对HMW-MAA的抗体和至少一种针对乳腺癌干细胞标记物的抗体。检测到HMW-MAA抗体和乳腺癌干细胞标记物的抗体的结合可确定存在HMA-MAA+乳腺癌干细胞。
就HMW-MAA而言，它是一种高度糖基化的膜内在硫酸软骨素。它由N-连接的280kDa的糖蛋白组分和450kDa的硫酸软骨素蛋白多糖组分构成。这两种组分共享相同的核心蛋白。通过使用小鼠和人单克隆抗体已鉴定出其许多抗原决定簇。它们在黑色素瘤细胞系上和在黑色素瘤损伤中显示出异源表达。HMW-MAA在黑色素瘤细胞的生长和转移潜能中起作用，尽管一项报告观察到HMW-MAA在乳腺癌细胞上表达(Dell′Erba等，(2001)Anticancer Res.03-04；21(2A)：925-30)，但目前发现HMW-MAA只在乳腺癌干细胞上表达，因为目前没有证据表明癌症干细胞也表达肿瘤细胞表达的抗原。结合这项发现，我们证明几乎所有获自乳腺癌患者胸腔积液的乳腺癌干细胞都包含表达HMW-MAA的乳腺癌干细胞。此外，我们证明本发明的方法可用于抑制动物模型中接种了我们已经确定表达HMW-MAA的人乳腺癌干细胞所形成癌的转移。再者，我们证明用本发明的方法可有效抑制由表达HMW-MAA的人乳腺癌干细胞所形成癌的切除后复发。因此，预计该方法通过寻靶表达HMW-MAA的乳腺癌干细胞而为乳腺癌患者提供了独特的疗法。
乳腺癌干细胞被认为是表达CD44(“CD44+”)但不表达CD24(“CD24-”)或者相对于正常细胞或非干细胞表达少量CD24(“CD241o”)的那些乳腺癌细胞。ESA也已知是乳腺癌干细胞的一种标记物，而B38.1已知是乳腺癌细胞。非干细胞被认为是表达CD2、CD3、CD10、CD16、CD18、CD31、CD45、CD64和CD140b中一种或多种的那些细胞。因此，表达CD2、CD3、CD10、CD16、CD18、CD31、CD45、CD64或CD140b中任一种的细胞不视作乳腺癌干细胞。本领域的技术人员将了解，用来鉴定乳腺癌干细胞的其他标记物可以是已知的或在之后被鉴定，并可与本发明联用来鉴定乳腺癌干细胞。
采用诸如免疫组织化学或细胞分选等常规方法，可将上述标记物用于鉴定乳腺癌干细胞。在一个实施方式中，采用Al-Hajj等(PNAS(2003)第100卷，3984-3983页)描述的细胞分选法和标记物可基本鉴定乳腺癌干细胞。本发明对该方法进行了改进从而能用抗-HMW-MAA抗体鉴定表达HMW-MAA的乳腺癌干细胞。
在一个实施方式中，可采用标准细胞分选程序通过流式细胞术来鉴定乳腺癌干细胞。例如，采用常规技术获自患者渗出液或活检的细胞可首先用菲可溶液(通常为500ML-2L)处理以除去细胞碎片和红血细胞污染。也可进行门控(Gating)(例如，用针对CD45的抗体)以区分血细胞。通过流式细胞染色进行乳腺癌干细胞表型分析可鉴定出“阴性谱系”细胞(对CD2、3、10、16、18、31、45、64、140b为阴性，例如，采用PE标记的抗体)。对于FACS分析，可利用CD44+-FITC标记的抗体/CD241o PerCP标记的抗体(所有抗体都来自加州圣何塞的BD/P公司(BD/Pharmingen，San Jose，CA))检测人乳腺癌患者胸腔积液的细胞。从恶性肿瘤渗出液中分选出的细胞可任选首先用抗-PE包被的珠处理以除去谱系标记物呈阳性的细胞从而大大减少非癌性干细胞的数量并因此缩短细胞分选时间。
在另一实施方式中，可通过流式细胞术分选ESA+CD44+CD24-/low细胞以评定患者样品中各种细胞群的存在和比例，如Al-Hajj等所述。与这种染色联合或相继地，ESA+CD44+CD24-/low细胞可用抗-HMW-MAA抗体染色以鉴定表达HMW-MAA的乳腺癌干细胞。任选地，可用一种以上针对HMW-MAA的抗体进行染色，所述抗体各自针对HMW-MAA的不同表位。适用于该方法的单克隆抗体的非限制性例子包括指定为225.28的抗-HMW-MAA抗体和/或指定为763.74的单克隆抗体。
本发明的HMW-MAA抗体可用于各种处理乳腺癌的诊断试验、成像法和治疗方法。例如，可在处理前后对获自个体的样品进行分析以确定本发明方法抑制个体中乳腺癌干细胞生长或将其清除的效率，所述分析方法例如对处理前后的活检样品进行分析、免疫组织化学分析、或细胞分选分析，以确定是否存在表达HMW-MAA的乳腺癌干细胞。
在涉及HMW-MAA+乳腺癌干细胞的诊断或治疗应用中，抗-HMW-MAA抗体可与各种部分偶联。例如，抗-HMW-MAA抗体可与治疗剂偶联以便能将治疗剂定位于表达HMW-MAA的乳腺癌干细胞。合适治疗剂的例子包括但不限于抗肿瘤药、毒素、放射性试剂、细胞因子、二抗或酶。细胞毒剂的例子包括但不限于蓖麻毒蛋白、蓖麻毒蛋白A-链、阿霉素、柔红霉素、紫杉酚、溴化乙锭、丝裂霉素等等。
在另一实施方式中，抗-HMW-MAA抗体可偶联于放射性试剂。有许多放射性同位素可与mAb偶联，从而能使表达HMW-MAA的乳腺癌干细胞成像或将其选择性破坏。为选择性破坏细胞，可将抗体偶联于高度放射性的原子，如In111、Ar211、I131、I125、Y90、Re186、Re188、Sm153、Bi212、P32、Pb212和Lu的放射性同位素。
当用抗体偶联物来鉴定表达HMW-MAA的乳腺癌干细胞时，所述抗体偶联物可包含任何合适的可检测的标记物，所述标记物包括但不限于放射性同位素、荧光化合物、生物发光化合物、化学发光化合物、金属螯合剂或酶。例如，某些放射性同位素可用于闪烁研究，如Tc99m(亚稳态锝-99)、I123，或用于核磁共振(NMR)成像(也称为磁共振成像，或“MRI”)的自旋标记的原子，如I123、I131、I124、F19、C13、N15、O17或钆(Gadlinium)(III)或锰(II)。可采用已知方法将这种标记掺入抗体内。“免疫闪烁法中的单克隆抗体”(Monoclonal Antibodies inImmunoscintigraphy)(Chatal，CRC出版社(CRC Press)，1989)详细描述了适用的方法。
除了本文所述的抗体，也可制造其他HMW-MAA抗体。制造单克隆和多克隆抗血清的方法是本领域熟知的。抗体或片段也可通过重组方法制造。或者，也可通过诸如噬菌体展示和转基因方法等方法来制造完整的人单克隆抗体(Vaughan等，1998，Nature Biotechnology 16：535-539)。例如，可用人Ig基因大组合文库来产生完整的人抗-HMW-MA单克隆抗体(即噬菌体展示)；(Griffiths和Hoogenboom，“建立体内免疫系统：来自噬菌体展示文库的人抗体”(Building an in vitro immune system：human antibodies from phage displaylibraries)，选自Clark，M.编的《通过蛋白质工程构建用于人类预防和治疗用途的抗体分子》(Protein Engineering of Antibody Molecules for Prophylactic andTherapeutic Applications in Man)，诺丁汉学术出版社(Nottingham Academic)，45-64页(1993)；Burton和Barbas，“来自组合文库的人抗体”(Human Antibodiesfrom combinatorial libraries)，同上，65-82页)。
可通过任何合适途径给予抗-HMW-MAA抗体，所述途径包括肠胃外、皮下、腹膜内、肺内和鼻内。肠胃外输注包括肌内、静脉内、动脉内、腹膜内、淋巴管内或皮下给药。此外，可通过脉冲输注(pulse infusion)给予抗体，例如采用递减剂量的抗体。
也可与抗-HMW-MAA抗体一起给予其他化合物，如化疗剂、免疫抑制剂和/或细胞因子。组合给药可包括采用不同制剂或单一药物制剂的同时给药，也可包括以任何顺序的连续给药，优选存在两种(或所有)活性剂同时发挥其生物活性的时期。
可与已知的药学上可接受的载体、赋形剂或稳定剂混合来制备包含抗-HMW-MAA抗体的治疗制剂。本领域技术人员将了解，药学上可接受的载体或稀释剂的形式和特性由要与其混合的活性成分的用量、给药途径和其他熟知的因素，如个体大小和疾病阶段决定。
提供以下示例性实施例来进一步描述而非限制本发明。
实施例1
该实施例证实，乳腺癌干细胞系的某乳腺癌干细胞亚群表达HMW-MAA。
用HMW-MAA-特异性mAb 763.74、TP61.5和VF1-TP41.2染色7种人乳腺癌细胞系(图1A和1B)，证明细胞系MDA-MB-435中至少80％的CD44+、CD24lo细胞被HMW-MAA-特异性mAb染色，细胞系MDA-MB-231和HS578T中分别有约70％和50％被染色，而细胞系MCF-7和SUM-149有不到4％被染色。值得注意的是，被三种HMW-MAA-特异性mAb染色的CD44+、CD24lo细胞的比例在多次细胞培养传代后稳定，这说明乳腺癌干细胞表达HMW-MAA是一种稳定的特征。
实施例2
该实施例证实乳腺癌干细胞表达的HMW-MAA的分子特征。为表征HMW-MAA-特异性mAb染色乳腺癌干细胞的分子基础，在Western印迹中用mAb 763.74检测人乳腺癌细胞系MDA-MB-435的裂解物。具体地说，如图2所示，用8％SDS-聚丙烯酰胺凝胶分离CD44+CD24lo乳腺癌细胞MDA-MB-435的裂解物以用HMW-MAA-特异性mAb 763.74(泳道3)和同种型对照mAbMK2-23(泳道6)进行免疫印迹分析。不表达HMW-MAA的人黑色素瘤细胞M14(泳道1和4)以及在HMW-MAA cDNA转染后表达HMW-MAA的M14/HMW细胞(泳道2和5)用作对照。如图2所示，鉴定出HMW-MAA中有两种特征性组分。
实施例3
该实施例证实人乳腺癌细胞系MDA-MB-435s中的CD44+/CD24-/low乳腺癌干细胞表达HMW-MAA。如图3A所示，用CD24-、CD44-特异性mAb染色MDA-MB-435s细胞显示＞80％的细胞为所示的CD44+/CD24-/low乳腺癌干细胞。如图3B所示，用HMW-MAA-特异性mAb 225.28(下图)和同种型对照mAb(上图)染色假定的CD44+/CD24-/low乳腺癌干细胞显示99.1％的CSC为HMW-MAA阳性。因此证实人乳腺癌干细胞系表达HMW-MAA。
实施例4
该实施例证实HMW-MAA-特异性mAb 763.74和225.28在能抑制SCID小鼠中人乳腺癌干细胞(MDA-MB-435s)的肺转移。结果示于图4。为获得图4所示的数据，在第0天给每只SCID小鼠静脉内注射人乳腺癌细胞MDA-MB-435s(2×106)。随后将所有具有肿瘤的小鼠随机分成三组(5只/组)。从第3天开始给其中一组腹膜内注射HMW-MAA-特异性mAb 763.74，给另一组注射HMW-MAA-特异性mAb 225.28(100μg/小鼠)，每周两次，总共注射9次。第三组小鼠注射同种型对照抗体。在第34天对所有小鼠实施安乐死并对肺转移结节进行计数。HMW-MAA-特异性mAb处理组和同种型对照抗体处理组之间的差异显著(p＜0.001)。
因此，该实施例证实，给予两种不同HMW-MAA-特异性mAb中的任何一种都可抑制通过接种表达HMW-MAA的人乳腺癌干细胞在动物模型中产生的肿瘤的转移，而给予不结合HMW-MAA的同种型对照mAb对于抑制这种转移无效。
实施例5
该实施例证实用mAb 225.28能抑制人乳腺癌干细胞的术后肺转移。为获得图5所示的数据，进行以下方案：
第0天：皮下接种乳房脂肪瘤；第7天：以200μg/小鼠进行mAb 225.28处理，每周两次；第71天：手术除去肿瘤；第103天：停止处理；第134天：杀死小鼠并收集肺以供转移分析。
由图5可见，给予mAb 225.28导致在除去用表达HMW-MAA的人乳腺癌干细胞接种动物模型获得的肿瘤后在统计学上显著抑制肺转移。
实施例6
该实施例证实针对HMW-MAA的mAb能抑制人乳腺癌的术后复发。
为获得表1所示数据，在第0天将人乳腺癌干细胞MDA-MB-435s(2×106)注射入每只SCID小鼠的乳房脂肪垫。随后将所有具有肿瘤的小鼠随机分成三组(5只/组)。从第7天开始给其中一组腹膜内注射HMW-MAA-特异性mAb763.74，给另一组注射HMW-MAA-特异性mAb 225.28(200μg/小鼠)，每周两次，总共注射18次。第三组小鼠注射同种型对照抗体。在第71天通过手术除去小鼠的所有肿瘤。以相同的方案继续用mAb处理，但只注射9次。在第131天杀死所有小鼠，检测并分析局部肿瘤复发和肺转移。
表1
| mAb | 225.28 | F3C25 | 763.74 |
| 肿瘤复发数/组 | 0/5 | 3/5(1只死亡) | 1/5 |
由表1可见，给予两种不同的HMW-MAA-特异性mAb中的任何一种都能抑制用表达HMW-MAA的人乳腺癌干细胞接种动物模型获得的肿瘤的复发，而识别不相关抗原的同种型对照(F3C25)不能抑制这种复发。
实施例7
该实施例证实乳腺癌患者胸膜渗出液中的乳腺癌干细胞亚群表达HMW-MAA。
为获得表2总结的数据，先后用抗-HMW-MAAmAb(克隆225.28、763.74、TP41.2或TP61.5)和PE-标记的抗-小鼠IgG标记乳腺癌患者的胸腔积液细胞。洗涤之后用FITC-标记的抗-CD24、PerCP-标记的抗-CD45、APC-标记的抗-CD44和7-AAD染色细胞。通过流式细胞术分析CD45-7AAD-细胞或CD45-7AAD-HMW-MAA+细胞中CD44+CD24-群体的百分比。将CD45-7AAD-HMW-MAA+群体中CD44+CD24-细胞的百分比除以CD45-7AAD-群体中CD44+CD24-细胞的百分比来计算通过门控HMW-MAA阳性细胞的CD44+CD24-群体富集情况，结果示于每格的括号中。各患者样品的最高富集倍数显示在右栏。
表2
因此，该实施例证实人乳腺癌患者中存在表达HMW-MAA的乳腺癌干细胞。
已经通过上述实施例描述了本发明。本领域技术人员显然可对上述方法和组合物进行常规改进，这种改进属于随附权利要求书的范围内。
Claims (20)
1.一种抑制个体的乳腺癌干细胞生长的方法，所述方法包括给予该个体有效量的与高分子量-黑色素瘤相关抗原(HMW-MAA)反应的抗体或其HMW-MAA反应性片段，其中所述乳腺癌干细胞表达HMW-MAA+。
2.如权利要求1所述的方法，其特征在于，所述抗体是单克隆抗体。
3.如权利要求1所述的方法，其特征在于，所述片段选自Fab、Fab’、F(ab’)2或Fv。
4.如权利要求2所述的方法，其特征在于，所述单克隆抗体与选自毒素或放射性同位素的试剂偶联。
5.如权利要求4所述的方法，其特征在于，所述放射性同位素选自I123、I125、I124或I131。
6.如权利要求1所述的方法，其特征在于，所述抗体与化疗剂同时或相继给予。
7.如权利要求1所述的方法，其特征在于，所述抗体通过选自下组的途径给予：肠胃外、皮下、腹膜内、静脉内、淋巴管内和肺内给药。
8.如权利要求1所述的方法，其特征在于，所述抗体在切除乳腺癌之后给予。
9.一种抑制个体的乳腺癌转移的方法，其中所述乳腺癌包含HMW-MAA+乳腺癌干细胞，所述方法包括给予该个体有效量的与HMW-MAA反应的抗体或其HMW-MAA反应性片段。
10.如权利要求9所述的方法，其特征在于，所述抗体是单克隆抗体。
11.如权利要求9所述的方法，其特征在于，所述片段选自Fab、Fab’、F(ab’)2或Fv。
12.如权利要求10所述的方法，其特征在于，所述单克隆抗体与选自毒素或放射性同位素的试剂偶联。
13.如权利要求12所述的方法，其特征在于，所述放射性同位素选自I123、I125、I124或I131。
14.如权利要求9所述的方法，其特征在于，所述抗体与化疗剂同时或相继给予。
15.如权利要求9所述的方法，其特征在于，所述抗体通过选自下组的途径给予：肠胃外、皮下、腹膜内、静脉内、淋巴管内和肺内给药。
16.如权利要求9所述的方法，其特征在于，所述抗体在切除乳腺癌之后给予。
17.一种检测HMA-MAA+乳腺癌干细胞的方法，所述方法包括将抗体组合物给予个体或使之与获自该个体的生物样品接触，所述组合物包含针对HMW-MAA的抗体或至少一种针对乳腺癌干细胞标记物的抗体，其中同时检测到HMW-MAA抗体和至少一种乳腺癌干细胞标记物的结合可确定存在HMA-MAA+乳腺癌干细胞。
18.如权利要求17所述的方法，其特征在于，所述抗体是单克隆抗体。
19.如权利要求18所述的方法，其特征在于，所述单克隆抗体与放射性同位素偶联。
20.如权利要求19所述的方法，其特征在于，所述针对乳腺癌干细胞标记物的抗体针对CD44、CD24及其组合。
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