Chinese Journal of Tissue Engineering Research
Previous Articles Next Articles
Li Qiang1, Liu Zhen-shan2
Received:
2012-05-07
Revised:
2012-06-19
Online:
2013-03-05
Published:
2013-03-05
Contact:
Liu Zhen-shan, Attending physician, Dezhou Fire detachment of Chinese Armed Police Force, Dezhou 253000, Shandong Province, China bubbledouble@163.com
About author:
Li Qiang, Attending physician, Department of Hematology, People’s Hospital of Liaocheng, Liaocheng 252002, Shandong Province, China
iama31415926@sina.com
CLC Number:
Li Qiang, Liu Zhen-shan. Source and differentiation of multiple myeloma stem cells[J]. Chinese Journal of Tissue Engineering Research, doi: 10.3969/j.issn.2095-4344.2013.10.028.
2.1 肿瘤干细胞 肿瘤组织内细胞可分为两群,一部分细胞组成肿瘤组织的主要细胞,一部分细胞数量很少,但可决定肿瘤的发生、发展和转移,这些细胞称为肿瘤干细胞。2006年,AACR对肿瘤干细胞的定义是具有干细胞特性(自我更新和分化)的肿瘤细胞,其对肿瘤干细胞的定义是功能性的,只要满足了干细胞的更能特点即可成为肿瘤干细胞[1]。 2.1.1 来源和鉴定 目前已在乳腺、脑、胰腺、前列腺、结肠、白血病和骨髓瘤等肿瘤组织中发现了肿瘤干细胞。肿瘤干细胞具有自我更新、分化和无限增殖3种生物学特性,可以产生维持肿瘤组织的肿瘤干细胞和分化的肿瘤细胞群,对常规化疗有抵抗作用[2-4]。由于机体内正常干细胞也具有这3种生物学特性,因此认为肿瘤干细胞来源于机体的正常干细胞。但也有研究表明肿瘤干细胞来源于突变的祖细胞[5]。大多数肿瘤干细胞首次分离时都是基于某种表面标志,是分离肿瘤干细胞的常用方法,作为目前的最佳选择。这种方法的优点是,只要明确了肿瘤干细胞的表面标志就能有效分离肿瘤干细胞[6-8]。肿瘤干细胞的来源还不清楚,在肿瘤发展过程中会不断发生突变,分子标志可能会有所改变。侧群细胞筛选法广泛用于肿瘤干细胞的分选。相对静止的肿瘤干细胞很少吸收荧光染料 Hoechst 33342或者细胞的ATP结合盒转运体能够泵出Hoechst 33342,利用此特性可以利用流式细胞术的方法筛选肿瘤干细胞。 2.1.2 信号通路 现在认为肿瘤不断生长的另一个重要原因是干细胞自我更新调节机制中的某些基因发生紊乱,使得肿瘤组织中那些具有自我更新能力的肿瘤细胞的数量不断增加,肿瘤组织能够不断的增大。肿瘤干细胞在信号转导通路方面与肿瘤细胞有很大不同。多种与细胞发育和增殖相关的信号通路在肿瘤干细胞中持续激活。已有的研究发现,Notch(脑、乳腺)、Wnt(脑、乳腺、肝细胞癌)、Shh(脑、多发性骨髓瘤)、核因子κB(白血病、乳腺)、PI3K (白血病、乳腺、肝)、PTEN/ mTOR(白血病、乳腺)、STAT3(乳腺、肝细胞癌)、Ras/MAPK(乳腺)、骨形态发生蛋白(脑)、转化生长因子β(乳腺)和Bmi1(白血病乳腺、脑)等信号通路可在肿瘤干细胞中异常激活。维持肿瘤干细胞的信号通路中,调节自我更新的发育信号通路(Notch、Wnt、Shh)对肿瘤干细胞尤为重要。Guzman等[9]发现AML细胞核因子κB通路持续激活,核因子κB通路特异性抑制剂能够诱导白血病干细胞凋亡,但对正常干细胞无影响。核因子κB通路对乳腺癌干细胞也非常重要,特异性抑制剂可以抑制乳腺癌干细胞增殖[10]。PI3K信号通路可维持白血病、乳腺癌和肝癌肿瘤干细胞生物学特性,其上下游蛋白PTEN和mTOR在白血病发病中起重要作用[11]。STAT3、Ras/MAPK、骨形态发生蛋白、转化生长因子β和Bmi1信号通路在维持肿瘤干细胞特性方面也有重要作用。 2.2 多发性骨髓瘤干细胞表面标记 Hamburger等[12]第1次使用软琼脂,体外培养出了骨髓瘤细胞克隆,骨髓瘤克隆形成率约为0.001%-0.1%。尽管多发性骨髓瘤表现为浆细胞增生,但这些细胞缺乏明显的增殖能力,而具有B细胞相似表型的细胞却具有克隆形成能力[13-14]。接触抗原后,V(D)J基因重排后的成熟B细胞,进入生发中心,免疫球蛋白发生类别转化和体细胞高频突变,多发性骨髓瘤浆细胞免疫球蛋白基因序列是体细胞高频突变的,在疾病过程中序列保持不变,说明多发性骨髓瘤是发生于生发中心B细胞以后的疾病[15]。 Kirshner等[16]利用新的3D细胞培养系统培养多发性骨髓瘤细胞发现,表达B细胞标志的细胞能够形成克隆。Huff等[17]体外比较了CD138-和CD138+细胞对4种药物(地塞米松、来那度胺、硼替佐米和羟基环磷酰胺)的敏感性,发现这几种药物均可以抑制CD138+的恶性浆细胞生长,但对CD138-的浆细胞无此作用。后来的实验中他们发现CD138-细胞表达记忆B细胞标志CD27和B细胞标志CD20的细胞具有体外致瘤性,在NOD/SCID小鼠体内进行初次和二次移植后均能引起与原发肿瘤类型相同的多发性骨髓瘤的发生。B细胞抗体利托昔单抗体外实验中限制克隆形成,利托昔单抗治疗多发性骨髓瘤的临床试验中,表现出一定治疗效果[18]。骨髓移植中产生的移植物抗宿主反应可以获得长期的缓解,可能与免疫为基础的治疗方法清除了骨髓瘤干细胞有关。以上研究充分证明记忆B细胞在多发性骨髓瘤中扮演了肿瘤干细胞的角色。 Pilarski 等[19]将粒细胞集落刺激因子动员的外周血细胞进行心内或直接骨内注射入NOD/SCID小鼠,可以形成溶骨性破坏和恶性浆细胞瘤,因为细胞中不含有浆细胞,认为肿瘤形成起源于B细胞。其后又从骨髓瘤患者分离出克隆性B细胞,接种于NOD/SCID小鼠体内,可以成功移植,形成肿瘤。记忆B细胞具有自我更新能力,维持长期的抗原识别功能,记忆B细胞高表达造血和胚胎干细胞自我更新的基因。能够形成克隆的细胞表达B细胞的表面标志CD45、CD19、CD20和CD22,克隆形成B细胞表型类似记忆B细胞,而记忆B细胞具有自我更新能力,维持长期的抗原识别功能,说明骨髓瘤起源于记忆B细胞。克隆形成骨髓瘤的CD138-细胞表达记忆B细胞抗原标志CD19+、CD27+,CD19+CD27+CD138-细胞能在NOD/SCID鼠上形成骨髓瘤,产生CD138+浆细胞,而且分离的CD19+细胞能够再次形成肿瘤,因此克隆形成骨髓瘤细胞是记忆B细胞[20]。 目前,对多发性骨髓瘤肿瘤干细胞的分子标志还有争议。Yaccoby等[21]用人胚胎骨片段移植于SCID小鼠,形成人源化的SCID小鼠,成熟的CD38+ CD45-的浆细胞能够诱导发病,产生循环M蛋白、高钙血症和人骨骼吸收。而CD38-CD45+的外周B细胞不能移植,说明仅成熟浆细胞就可成瘤。各研究组报告的肿瘤干细胞生物学特性不同可能与功能分析的方法不同有关。尽管干细胞的自我更新能力是内在特性,但干细胞龛内的因子也能够调控干细胞的增殖过程[22]。因此,各实验中的外部因子的不同,可能影响肿瘤干细胞的定植、存活和增殖。例如Yaccoby 等[21]利用人胚胎骨片段形成人源化的SCID小鼠,可能利于浆细胞增殖,但缺乏B细胞存活的因子。NOD/SCID小鼠骨髓开始可能不适于成熟浆细胞增殖,但移植B细胞后,可能诱导骨髓改变,适于浆细胞存活。另外,注射位点、细胞运输和细胞分选方法可能也会影响实验结果。多发性骨髓瘤的分期和治疗都会影响肿瘤干细胞的致瘤性,而且多发性骨髓瘤可能包含多种不同的疾病,有不同的致瘤性。 2.3 多发性骨髓瘤干细胞信号通路 多发性骨髓瘤干细胞通过与正常干细胞一样的途径和信号分子如Hedgehog,Wnt及Notch等而具有自我更新的特性,当这些信号传导通路出现异常时,就导致了肿瘤的发生及肿瘤细胞的无控制性增长和扩增。 2.3.1 Hedgehog信号通路 通常Hedgehog信号通路在缺乏配体时处于关闭状态,跨膜蛋白受体Ptch抑制Smo的活性,导致Hedgehog目标基因的转录活性受到抑制而失活,当该通路组分发生突变则导致肿瘤的发生[23-25]。Hedgehog信号通路参与多种癌症的发生并在多发性骨髓瘤的肿瘤干细胞中处于活化状态。已证实多发性骨髓瘤细胞系和肿瘤样本存在克隆性极强的CD19+CD138-肿瘤干细胞,许多患者肿瘤组织CD19+CD138-肿瘤干细胞Smo mRNA水平显著上调。在骨髓瘤细胞系NCI-H929中,肿瘤干细胞(CD138-)Smo表达水平也非常高。采用cyclopamine或单克隆抗体5E1抑制Hedgehog通路,其克隆形成能力明显受阻,实验表明在多发性骨髓瘤干细胞中,如果Ptch下调,而Smo高表达,将导致Hedgehog通路激活而显著扩增多发性骨髓瘤胞干细胞数量。 2.3.2 Wnt信号通路 Wnt信号通路活化后,可激活肿瘤发生和转移中起重要作用的靶基因(如原癌基因c-myc、细胞周期蛋白D1和金属蛋白酶等)的转录和表达,Wnt通路正是结合了c-myc来实现对干细胞的增殖与分化的调控。研究表明此信号通路能通过扩增干细胞数量而导致肿瘤的发生。目前国外有研究已证实了多种多发性骨髓瘤细胞系及多发性骨髓瘤患者骨髓中Wnt信号通路是显著激活,可检测到β连环蛋白(β-catenin)过度表达,且在体外证实了Wnt/β-catenin信号通路有调节骨髓瘤细胞增殖和侵袭的作用[26]。有实验证明蛋白β连环蛋白的表达水平在复发/难治组较初治组明显增高,提示β连环蛋白的高表达可能与多发性骨髓瘤进展或细胞耐药有关,β连环蛋白可能可作为逆转骨髓瘤细胞耐药治疗的新靶点。 2.3.3 Notch信号通路 Notch信号分子是一个跨膜系统,大量转基因动物研究证明了Notch信号通路在干细胞和早期祖细胞的自我更新中发挥作用,活化的Notch信号传导途径可以调控其增殖分化[27-31]。Notch信号通路在干细胞中的作用的研究不如Wnt信号通路多,但研究显示Notch信号在造血干细胞自我更新的调控中发挥着重要作用,Notch信号的阻断导致体外造血干细胞分化的加速和体内造血干细胞的减少。实验检测了Notch信号被阻断的造血干细胞重建造血系统的能力,结果提示Notch信号的阻断导致了更高的分化率,从而阻断了造血干细胞未分化状态的维持。所以认为活化的Notch信号能抑制造血干细胞分化,维持其多潜能性,并可进一步促进其增殖。有实验表明,Notch受体及配体在多种肿瘤细胞及肿瘤衍生的细胞系中均有表达,动物实验也进一步确认Notch信号的过度活化将导致肿瘤的发生,目前已在多种肿瘤中发现Notch信号通路的改变能促进干细胞的自我更新及阻断分化,其在骨髓瘤干细胞中的表达有待进一步研究。"
[1] Clarke MF, Dick JE, Dirks PB, et al. Cancer stem cells- perspectives on current status and future directions: AACR Workshop on cancer stem cells. Cancer Res. 2006;66(19): 9339-9344.[2] 李青,唐良萏,汪艳,等.人卵巢肿瘤细胞系3AO中肿瘤干细胞的分离鉴定[J].华中科技大学学报:医学版,2012,41(1):36-40.[3] 侯萍,李剑平.肿瘤干细胞的研究进展[J].中国组织工程研究与临床康复, 2011, 15(14):2629-2632.[4] 凌斌,陈静,孙洁.肿瘤干细胞与干细胞:来源、分化及其相关性[J]. 中国组织工程研究与临床康复, 2009, 13(49): 9743-9746.[5] Krivtsov AV, Twomey D, Feng Z, et al. Transformation from committed progenitor to leukaemia stem cell initiated by MLL-AF9. Nature.2006;442(7104):818-822.[6] Com?a S, Ciuculescu F, Raica M. Mesenchymal stem cell-tumor cell cooperation in breast cancer vasculogenesis. Mol Med Report.2012;5(5):1175-1180.[7] Estrada-Bernal A, Palanichamy K, Ray Chaudhury A, et al. Induction of brain tumor stem cell apoptosis by FTY720: a potential therapeutic agent for glioblastoma. Neuro Oncol. 2012;14(4):405-415.[8] Choijamts B, Jimi S, Kondo T, et al. CD133+ cancer stem cell-like cells derived from uterine carcinosarcoma (malignant mixed Müllerian tumor). Stem Cells. 2011;29(10):1485-1495.[9] Guzman ML, Rossi RM, Neelakantan S, et al. An orally bioavailable parthenolide analog selectively eradicates acute myelogenous leukemia stem and progenitor cells. Blood. 2007;110:4427-4435.[10] Zhou J, Zhang H, Gu P, et al. NF-kappaB pathway inhibitors preferentially inhibit breast cancer stem-like cells. Breast Cancer Res Treat. 2008;111(3):419-427.[11] Zhou J, Wulfkuhle J, Zhang H, et al. Activation of the PTEN/mTOR/STAT3 pathway in breast cancer stem-like cells is required for viability and maintenance. Proc Natl Acad Sci.2007;104:16158-16163.[12] Hamburger AW, Salmon SE. Primary bioassay of human tumor stem cells. Science. 1977;197(4302):461-463.[13] Matsui W, Wang Q, Barber JP et al. Clonogenic Multiple Myeloma Progenitors, Stem Cell Properties, and Drug Resistance. Cancer Res.2008;68(1):190-197.[14] Matsui W, Huff CA, Wang Q, et al. Characterization of clonogenic multiple myeloma cells. Blood.2004;103(6): 2332-2336.[15] Brennan SK, Matsui W. Cancer stem cells: controversies in multiple myeloma. J Mol Med.2009;87(11):1079-1085.[16] Kirshner J, Thulien KJ, Martin LD, et al. A unique three-dimensional model for evaluating the impact of therapy on multiple myeloma. Blood.2008; 112(7):2935-2945.[17] Huff CA, Matsui W. Multiple myeloma cancer stem cells. J Clin Oncol.2008; 26(17):2895-2900.[18] Treon SP, Pilarski LM, Belch AR, et al. CD20-directed serotherapy in patients with multiple myeloma: biologic considerations and therapeutic applications. J Immunother. 2002;25(1):72-81.[19] Pilarski LM, Hipperson G, Seeberger K, et al. Myeloma progenitors in the blood of patients with aggressive or minimal disease engraftment and self-renewal of primary human myeloma in the bone marrow of NOD SCID mice. Blood.2000; 95(3):1056-1065.[20] Ghosh N, Matsui W. Cancer stem cells in multiple myeloma. Cancer Lett.2009; 277(1):1-7.[21] Yaccoby S, Epstein J. The proliferative potential of myeloma plasma cells manifest in the SCID-hu host. Blood.1999; 94(10):3576-3582.[22] Scadden DT. The stem-cell niche as an entity of action. Nature.2006; 441(7097):1075-1079.[23] Gallet A. Hedgehog morphogen: from secretion to reception. Trends Cell Biol, 2011;21(4):238-246.[24] Takebe N, Harris PJ, Warren RQ,et al.Targeting cancer stem cells by inhibiting Wnt, Notch, and Hedgehog pathways. Nat Rev Clin Oncol.2011;8(2):97-106.[25] Jagani Z, Dorsch M, Warmuth M. Hedgehog pathway activation in chronic myeloid leukemia. Cell Cycle.2010; 9(17):3449-3456.[26] Kim Y, Reifenberger G, Lu D, et al. Influencing the Wnt signaling pathway in multiple myeloma. Anticancer Res. 2011;31(2):725-730.[27] Klinakis A, Lobry C, Abdel-Wahab O, et al. A novel tumour-suppressor function for the Notch pathway in myeloid leukaemia. Nature.2011;473(7346):230-233.[28] Ranganathan P, Weaver KL, Capobianco AJ. Notch signalling in solid tumours: a little bit of everything but not all the time. Nat Rev Cancer.2011;11(5):338-351.[29] Wang Z, Ahmad A, Li Y, et al. Targeting notch to eradicate pancreatic cancer stem cells for cancer therapy. Anticancer Res.2011;31(4):1105-1113.[30] Galluzzo P, Bocchetta M. Notch signaling in lung cancer. Expert Rev Anticancer Ther.2011;11(4):533-540.[31] Bridges E, Oon CE, Harris A. Notch regulation of tumor angiogenesis. Future Oncol.2011;7(4):569-588. |
[1] | Pu Rui, Chen Ziyang, Yuan Lingyan. Characteristics and effects of exosomes from different cell sources in cardioprotection [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(在线): 1-. |
[2] | Zhang Xiumei, Zhai Yunkai, Zhao Jie, Zhao Meng. Research hotspots of organoid models in recent 10 years: a search in domestic and foreign databases [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(8): 1249-1255. |
[3] | Wang Zhengdong, Huang Na, Chen Jingxian, Zheng Zuobing, Hu Xinyu, Li Mei, Su Xiao, Su Xuesen, Yan Nan. Inhibitory effects of sodium butyrate on microglial activation and expression of inflammatory factors induced by fluorosis [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1075-1080. |
[4] | Wang Xianyao, Guan Yalin, Liu Zhongshan. Strategies for improving the therapeutic efficacy of mesenchymal stem cells in the treatment of nonhealing wounds [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1081-1087. |
[5] | Liao Chengcheng, An Jiaxing, Tan Zhangxue, Wang Qian, Liu Jianguo. Therapeutic target and application prospects of oral squamous cell carcinoma stem cells [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1096-1103. |
[6] | Xie Wenjia, Xia Tianjiao, Zhou Qingyun, Liu Yujia, Gu Xiaoping. Role of microglia-mediated neuronal injury in neurodegenerative diseases [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1109-1115. |
[7] | Li Shanshan, Guo Xiaoxiao, You Ran, Yang Xiufen, Zhao Lu, Chen Xi, Wang Yanling. Photoreceptor cell replacement therapy for retinal degeneration diseases [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1116-1121. |
[8] | Jiao Hui, Zhang Yining, Song Yuqing, Lin Yu, Wang Xiuli. Advances in research and application of breast cancer organoids [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1122-1128. |
[9] | Wang Shiqi, Zhang Jinsheng. Effects of Chinese medicine on proliferation, differentiation and aging of bone marrow mesenchymal stem cells regulating ischemia-hypoxia microenvironment [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1129-1134. |
[10] | Zeng Yanhua, Hao Yanlei. In vitro culture and purification of Schwann cells: a systematic review [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1135-1141. |
[11] | Kong Desheng, He Jingjing, Feng Baofeng, Guo Ruiyun, Asiamah Ernest Amponsah, Lü Fei, Zhang Shuhan, Zhang Xiaolin, Ma Jun, Cui Huixian. Efficacy of mesenchymal stem cells in the spinal cord injury of large animal models: a meta-analysis [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1142-1148. |
[12] | Hou Jingying, Yu Menglei, Guo Tianzhu, Long Huibao, Wu Hao. Hypoxia preconditioning promotes bone marrow mesenchymal stem cells survival and vascularization through the activation of HIF-1α/MALAT1/VEGFA pathway [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 985-990. |
[13] | Shi Yangyang, Qin Yingfei, Wu Fuling, He Xiao, Zhang Xuejing. Pretreatment of placental mesenchymal stem cells to prevent bronchiolitis in mice [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 991-995. |
[14] | Liang Xueqi, Guo Lijiao, Chen Hejie, Wu Jie, Sun Yaqi, Xing Zhikun, Zou Hailiang, Chen Xueling, Wu Xiangwei. Alveolar echinococcosis protoscolices inhibits the differentiation of bone marrow mesenchymal stem cells into fibroblasts [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 996-1001. |
[15] | Fan Quanbao, Luo Huina, Wang Bingyun, Chen Shengfeng, Cui Lianxu, Jiang Wenkang, Zhao Mingming, Wang Jingjing, Luo Dongzhang, Chen Zhisheng, Bai Yinshan, Liu Canying, Zhang Hui. Biological characteristics of canine adipose-derived mesenchymal stem cells cultured in hypoxia [J]. Chinese Journal of Tissue Engineering Research, 2021, 25(7): 1002-1007. |
Viewed | ||||||
Full text |
|
|||||
Abstract |
|
|||||