胃癌单克隆细胞球和CD44＋及CD133＋亚群成瘤能力的比较

目的研究胃癌干细胞亚群的筛选方法,探讨建立胃癌干细胞稳定亚群的可行性。方法利用流式细胞仪从MKN45、MKN25、SGC7901细胞株和人胃癌组织中分选出不同CD44和CD133表型的亚群,比较不同亚群和无血清培养基培养的单克隆细胞球细胞在裸鼠移植瘤模型中的成瘤能力。结果不同细胞的CD44＋和CD133＋亚群在低数量级时几乎不具有裸鼠皮下成瘤能力,在高数量级接种时成瘤能力与母系细胞差异无统计学意义;单克隆细胞球细胞在各数量级下的成瘤能力、瘤体积和生瘤速度均显著高于母系细胞。结论与CD44和CD133等细胞表面标志物筛选法相比,单克隆细胞球细胞可更好的作为胃癌干细胞研究的细胞学基础。     

Prognostic value of cancer stem cell marker CD133 expression in non-small cell lung cancer: a systematic review

Objective: To investigate the correlation between CD133-positive non-small cell lung cancer (NSCLC) and clinicopathological features and its impact on survival. Methods: A search in the Pubmed, Embase and Wanfang databases (up to July 15, 2013) was performed. Only articles in which CD133 antigen was detected in situ localization by immunohistochemical staining were included. This meta-analysis was done using RevMan 5.2 software. Outcomes included overall survival and various clinicopathological features. Results: A total of 1004 NSCLC patients from 11 studies were included. Meta-analysis showed that CD133 expression patients had a significant worse 5-year overall survival compared to the low expression ones (RR = 3.19, 95% CI: 2.05-4.98, P<0.0001 fixed random). With respect to clinicopathological features, CD133 expression by IHC method was closely correlated with tumor T stage (OR = 0.91, 95% CI: 0.59-1.39, P = 0.67 fixed-effect) and tumor grade (OR = 1.20, 95% CI: 0.80-1.79, P = 0.37 fixed-effect). Conclusion: CD133-positive NSCLC patients had worse prognosis, and was associated with common clinicopathological poor prognostic factors.     

CD133+胶质瘤干细胞在胶质瘤中的分布

恶性胶质瘤是人类最致命的肿瘤之一,因其血管丰富和弥漫浸润性生长,使得手术不能全部切除并极易复发,患者预后差.因胶质瘤干细胞(GSCs)是胶质瘤发生、发展的源头,因而越来越多的受到关注.干细胞标志物CD133广泛表达于人体多种肿瘤中,同时,它也足研究得最多且应用最广泛的胶质瘤干细胞表面标志物.了解CD133+胶质瘤干细胞在胶质瘤中的分布可为临床靶向治疗恶性胶质瘤提供依据.     

人鼻咽癌细胞株分选CD133~+细胞实验研究

目的观察CD133在人鼻咽癌细胞株中的表达,探讨CD133+肿瘤细胞的体外增殖及分化能力从而确定鼻咽癌肿瘤干细胞的标志。方法使用免疫细胞化学及流式细胞技术检测鼻咽癌CNE2Z细胞株中的表达,免疫磁珠分选技术纯化肿瘤细胞,体外培养,观察其增殖及分化能力。结果鼻咽癌CNE2Z细胞株中有0.168%呈微量阳性表达,利用免疫磁珠分选技术纯化细胞的百分比为87.2%,免疫磁珠富集的CD133+肿瘤细胞在无血清培养基中1、3、5、7天的吸光度均高于相同条件下未分选细胞和CD133-细胞;CD133+在培养体系中的比例逐日下降,至培养的第12天,由第1天的87.2%下降至0.1%。结论鼻咽癌CNE2Z细胞株中,CD133+癌细胞有比其他细胞亚群强的体外分化和增殖能力,具有自我更新、生成其他表型肿瘤细胞等干细胞样特性,CD133可能是肿瘤起始细胞的标志之一。     

低氧对结肠癌SW480细胞生存的影响及与细胞自噬的关系

 摘要：目的 观察低氧微环境对结肠癌细胞增殖、侵袭及自噬的影响,探讨低氧对结肠癌细胞生存及与自噬的关系。方法 免疫磁珠分选CD133+ 细胞;Boyden小室实验研究结肠癌细胞的侵袭;透射电子显微镜（TEM）观察细胞的超微结构;MDC荧光检测自噬囊泡;联合运用流式细胞仪和台盼蓝染色研究自噬与细胞增殖的关系。结果 Boyden小室实验中低氧微环境增加结肠癌细胞SW480进入下室的细胞数量;在实验组低氧的微环境中SW480细胞的MDC荧光强度由20.53±0.64降低至17.00±0.20,而CD133+ 细胞的MDC荧光强度由19.89±0.58增加至33.13±1.95（P<0.05）。台盼蓝染色实验显示,与对照组相比,低氧微环境中SW480活细胞率随着时间的延长而降低（P<0.05）;但CD133+ 活细胞率无显著改变。结论：低氧可增加SW480细胞的侵袭性,细胞自噬水平下降,活细胞率降低;CD133+细胞在低氧的微环境中自噬的发生增加,活细胞率基本不变。     

5氟尿嘧啶上调干细胞标记物CD133在结肠癌细胞中的表达

目的:研究通过5氟尿嘧啶(5-FU)对结肠癌干细胞标记物CD133表达的影响,探讨5-FU对结肠癌干细胞的影响。方法:用流式细胞仪检测CD133在结肠癌细胞株表面的表达,磁珠细胞分离的方法分离结肠癌细胞株DLD1中CD133阳性和阴性的细胞群,细胞克隆形成实验检测2群细胞的自我更新能力,新型四唑氮盐方法(MTS)检测2群细胞对5-FU敏感性的差异,qPCR方法检测5-FU处理结肠癌细胞后CD133mRNA水平的变化。结果:结肠癌细胞株DLD1、HT29、SW480、HCT116、Lovo、RKO细胞表面CD133的表达率分别为30.20%、82.00%、0.34%、91.80%、85.30%、0.28%。DLD1细胞中以CD133为标记有2群明显的细胞,MACS方法分离后阳性细胞群中CD133为87.21%±5.33%,而阴性细胞群中阴性细胞的比例为84.30%±4.65%;CD133阳性的细胞与未分离及CD133阴性细胞相比,克隆形成能力强(46.33%±4.44%vs31.00%±2.00%,P<0.05),对5-FU的敏感性下降20%,P<0.01。在DLD1和HT29细胞中,5-FU1mg/L上调CD133mRNA水平的表达,从1升为1.684±0.012(P<0.01)、HT29细胞从30.702±0.284升为49.379±0.460(P<0.01)。结论:与CD133阴性细胞相比CD133阳性细胞克隆形成能力强,对5-FU的敏感性下降;5-FU上调干细胞标记物CD133mRNA水平的表达,CD133阳性的结肠癌干细胞在5-FU的治疗过程中被富集。     

In Search of Liver Cancer Stem Cells

Recent research efforts in stem cell and cancer biology have put forth a “stem cell model of carcinogenesis” which stipulates that the capability to maintain tumor formation and growth specifically resides in a small population of cells called cancer stem cells. The stem cell-like characteristics of these cells, including their ability to self-renew and differentiate; and their limited number within the bulk of the tumor mass, are believed to account for their capability to escape conventional therapies. In the past few years, the hypothesis of stem cell-driven tumorigenesis in liver cancer has received substantial support from the recent ability to identify and isolate a subpopulation of liver cancer cells that is not only able to initiate tumor growth, but also serially establish themselves as tumor xenografts with high efficiency and consistency. In this review, stem cell biology that contributes to explain tumor development in the particular context of liver cancer will be discussed. We will begin by briefly considering the knowledge available on normal liver stem cells and their role in tissue renewal and regeneration. We will then summarize the current scientific knowledge of liver cancer stem cells, discuss their relevance to the diagnosis and treatment of the disease and consider the outstanding challenges and potential opportunities that lie ahead of us.     

侧群细胞及其在干细胞研究中的应用

肿瘤干细胞学说的提出为肿瘤的治疗提供了新的靶点和方向。尽管已经从白血病、乳腺癌、脑肿瘤等肿瘤中初步分离鉴定出肿瘤干细胞,但是还有许多恶性肿瘤由于肿瘤干细胞表面标记尚不清楚而无法鉴定,通过侧群细胞(side population cells,SP) 分选来初筛肿瘤干细胞,方法简便、有效,正被越来越多地应用。     

The utility and limitations of glycosylated human CD133 epitopes in defining cancer stem cells

Human CD133 (human prominin-1), a five transmembrane domain glycoprotein, was originally identified as a cell surface antigen present on CD34+ hematopoietic stem cells. Although the biological function of CD133 is not well understood, antibodies to CD133 epitopes have been widely used to purify hematopoietic stem and progenitor cells. The cancer stem cell (CSC) hypothesis postulates that a rare population of tumor cells possessing increased capacities for self-renewal and tumor initiation is responsible for maintaining the growth of neoplastic tissue. The expression of the CD133 epitopes, AC133 and AC141, has been shown to define a subpopulation of brain tumor cells with significantly increased capacity for tumor initiation in xenograft models. Following the discovery of the AC133/AC141+ population of brain tumor stem cells, the AC133 and AC141 epitopes have been extensively used as markers for purifying CSCs in other solid tumors. There are, however, several issues associated with the use of the AC133 and AC141 CD133 epitopes as markers for CSCs. The antibodies routinely used for purification of AC133 and AC141-positive cells target poorly characterized glycosylated epitopes of uncertain specificity. Discordant expression of the AC133 and AC141 epitopes has been observed, and the epitopes can be absent despite the presence of CD133 protein. In addition, CD133 expression has recently been shown to be modulated by oxygen levels. These factors, in combination with the uncertain biological role of CD133, suggest that the use of CD133 expression as a marker for CSCs should be critically evaluated in each new experimental system and highlight the need for additional CSC surface markers that are directly involved in maintaining CSC properties.     

CD133(+) human umbilical cord blood stem cells enhance angiogenesis in experimental chronic hepatic fibrosis

The in vivo angiogenic potential of transplanted human umbilical cord blood (UCB) CD133(+) stem cells in experimental chronic hepatic fibrosis induced by murine schistosomiasis was studied. Enriched cord blood-derived CD133(+) cells were cultured in primary medium for 3 weeks. Twenty-two weeks post-Schistosomiasis infection in mice, after reaching the chronic hepatic fibrotic stage, transplantation of stem cells was performed and mice were sacrificed 3 weeks later. Histopathology and electron microscopy showed an increase in newly formed blood vessels and a decrease in the fibrosis known for this stage of the disease. By immunohistochemical analysis the newly formed blood vessels showed positive expression of the human-specific angiogenic markers CD31, CD34 and von Willebrand factor. Few hepatocyte-like polygonal cells showed positive expression of human vascular endothelial growth factor and inducible nitric oxide synthase. The transplanted CD133(+) human stem cells primarily enhanced hepatic angiogenesis and neovascularization and contributed to repair in a paracrine manner by creating a permissive environment that enabled proliferation and survival of damaged cells rather than by direct differentiation to hepatocytes. A dual advantage of CD133(+) cell therapy in hepatic disease is suggested based on its capability of hematopoietic and endothelial differentiation.     

人CD133基因转染细胞株的建立及其生物学功能的初步研究

目的:构建人干细胞标记分子CD133-2转基因细胞并探讨CD133-2分子的生物学功能。方法:利用分段克隆和重叠PCR方法从人胎肝cDNA文库中克隆出人CD133-2全长基因,经双酶切后装入真核表达载体p IRES2-EGFP中,用脂质体法转染L929细胞,加入G418选择性培养基进行筛选。经RT-PCR、Western blot、流式细胞术(FCM)等方法鉴定转基因细胞;MTT法分析转基因细胞对T细胞的体外增殖作用;流式细胞仪分析T细胞表面活化分子标记CD4CD25、CD8CD25的表达。结果:成功克隆和构建了能稳定表达人CD133-2分子的转基因细胞CD133-2/L929,该转基因细胞与T细胞共培养可抑制其体外增殖,下调T细胞表面分子CD4CD25和CD8CD25的表达。结论:稳定表达CD133-2蛋白的转基因细胞株的建立为该基因功能的后续研究奠定了物质基础。     

肿瘤干细胞标志物ALDH1及CDl33在鼻咽癌中的表达及其临床意义

目的：研究乙醛脱氢酶1（aldehyde dehydrogenase 1,ALDHl）、CDl33在鼻咽癌组织中的表达情况,探讨其作为鼻咽癌干细胞标志物的临床病理意义。方法：应用免疫组织化学法检测112例鼻咽癌及20例正常组织中ALDHl和CDl33的表达情况。结果：鼻咽癌组织中ALDHl和CDl33表达高于正常组织（P〈0．05）。ALDHl表达在患者年龄、性别、病理分型、原发癌和转移癌、是否伴有颈部淋巴结转移等各组间差异均无统计学意义（均P〉0．05）。CDl33表达在鼻咽部原发癌与淋巴结转移癌之间差异有统计学意义（P〈0．01）,在患者年龄、性别、病理分型以及是否伴有颈部淋巴结转移等各组间差异均无统计学意义（均P〉0．05）。结论：ALDHl作为鼻咽癌肿瘤干细胞标志物有待进一步研究。CDl33可作为鼻咽癌干细胞较特异的表面标志物,在鼻咽癌转移方面可能起重要作用。