In vivo investigation of CD133 as a putative marker of cancer stem cells in Hep-2 cell line

Background

Mounting evidence suggests that most tumors consist of a heterogeneous population of cells with a subset population that has the exclusive tumorigenic ability. They are called cancer stem cells (CSCs). CSCs can self‐renew to generate additional CSCs and also differentiate to generate phenotypically diverse cancer cells with limited proliferative potential. They have been identified in a variety of tumors. In this study, we identify the marker of CSCs in the established human laryngeal tumor Hep‐2 cell line in vivo. Our in vitro experiment shown as CD133, a 5‐transmembrane glycoprotein expressed in Hep‐2 cell line. CD133 was supposed as a candidate of CSC in laryngeal carcinoma. In this study, the expression of CD133 was detected in a Hep‐2 cell line. Applying the magnetic cell sorting (MACS) technology, we reported the results of purifying CD133 positive cells from a Hep‐2 cell line. Three‐type cells' tumor‐forming ability was examined in vivo to identify the marker of CSCs in Hep‐2 cell line.

Methods

CD133 was selected as a putative marker of CSC in laryngeal carcinoma, Hep‐2 cell lines. Flow cytometry was used to detect the expression of CD133 in the Hep‐2 cell line. Immunomagnetic beads were applied to purify CD133‐positive cells. CD133(+), CD133(?) tumor cells, and unsorted Hep‐2 cells were injected into severe combined immune deficiency (SCID) mice individually to observe tumor‐forming ability.

Results

Only a small proportion (3.15% ± 0.83%) of cells in the Hep‐2 cell line express the CD133 marker. In comparison with CD133(?) tumor cells and unsorted cells, CD133(+) cells possess a marked capacity for tumor formation in vivo (p <.05).

Conclusion

CD133 is 1 of the markers for CSCs in human laryngeal tumors of the Hep‐2 cell line. Work on the characterization of these cells provides a powerful tool to investigate the tumorigenic process in the larynx and to develop therapies targeting the CSC. © 2008 Wiley Periodicals, Inc. Head Neck, 2009

     

人脑胶质瘤中肿瘤干细胞的分离、鉴定以及生物学性质研究

目的研究不同级别胶质瘤中肿瘤干细胞的构成,分布以及肿瘤干细胞的生长特性。方法取不同级别的脑胶质瘤细胞12例（WHOⅡ-Ⅳ级）,接种于含生长因子的无血清培养基中培养,观察细胞的生长特性;用单克隆生长试验确定肿瘤干细胞比例;检测脑肿瘤干细胞中CD133含量以及多项分化能力的表达情况。结果在Ⅱ-Ⅳ级别的人脑胶质瘤中,均发现有CD133阳性细胞的存在,其中有1.2%～13.9%的细胞具有单细胞增殖形成克隆球的能力。结论人脑胶质瘤组织中确实存在少数的脑肿瘤干细胞,有自我更新、多项分化以及致瘤能力。     

细胞株源人前列腺肿瘤干细胞的分选与鉴定

目的:论证人前列腺癌(prostate cancer,PCa)细胞株中是否存在干细胞亚群。方法:分别用免疫表型法和侧群(side population,SP)细胞法从5种人PCa细胞株(Du145、IA8、LNCaP、TSU-PrL和PC-3)中富集类干细胞,再应用软琼脂克隆形成试验初步验证类干细胞亚群的体外生长方式及成瘤能力。选择LNCaP源SP细胞(LNCaP/SP),依次采用免疫细胞化学技术、Transwell、MTT以及裸鼠致瘤试验,分别检测其干细胞标记物的表达情况、鉴定其体外增殖和侵袭能力以及动物体内的致瘤和转移潜能。结果:5种细胞株中均难以分选出免疫表型为CD133+CD44+的细胞亚群。除PC-3外,其余4株细胞可分选出呈现典型克隆性生长特点的SP细胞。体外克隆形成率在IA8、LNCaP和TSU-PrL源SP细胞与非侧群(non-side population,NSP)细胞间有显著性差异(P<0.05)。与LNCaP/NSP相比,LNCaP/SP的体外增殖和侵袭能力显著增强,同时阳性表达整合素α2、Nanog、CD44、OCT4以及ABCG2等5种干细胞标记物。而且,LNCaP/SP的皮下成瘤率、骨转移率及瘤体体积亦显著高于LNCaP/NSP(P<0.01)。结论:SP分选法更适合富集人PCa细胞株中类干细胞,LNCaP/SP细胞是PCa细胞株LNCaP中的肿瘤干细胞(cancer stem cell,CSC)。     

Progenitor cells are responsible for formation primary epithelial cultures in the prostate epithelial model

The adult stem cells (ASC) are supposed to regenerate epithelium. We hypothesized prostate epithelial CD133-positive ASC to be responsible for establishing the primary cell culture. The prostate epithelial stem cells were isolated using anti-CD133 microbeads in order to form different cell populations. The morphology of cultures developed from CD133⁺ and CD133⁻ prostate epithelial cells were compared with prostate epithelium cell culture obtained after simple isolation procedure. Four 8-week-old Wistar rats were used in the experiment and six cultures were obtained. Double CD133⁺ and CD133⁻ cultures from two rats were established after enzymatic digestion and positive selection by SuperMACS device, and two non-selected CD133⁺/CD133⁻ cultures were developed by simple prostate epithelial cell isolation from two other rats. The epithelial nature was confirmed by anti-cytokeratine antibodies. It was observed that growth of the CD133⁺/CD133⁻ and CD133⁺culture resembled epithelial-like prostate cell culture. It was not possible to establish epithelial-like culture from CD133⁻ cell population. The primary epithelial cell culture collapsed in a few days after the CD133-positive ASC were removed. We concluded that the epithelial progenitor cells are responsible for establishing primary prostate epithelial cultures in vitro.     

Role of microRNAs in the Regulation of Breast Cancer Stem Cells

There is increasing evidence that many human cancers, including breast cancer, are driven and maintained by cancer stem cells (CSCs) which mediate tumor metastasis and contribute to treatment resistance and relapse. Our group was the first to describe “breast cancer stem cells” (BCSCs) characterized by expression of the cell surface markers ESA and CD44 and the absence of expression of the marker CD24. More recently, we have demonstrated that breast cancer cells contain subpopulations with stem cell properties that can be isolated by virtue of their expression of Aldehyde dehydrogenase (ALDH) as assessed by the Aldefluor assay. Interestingly, these markers identify overlapping, but not identical cell populations. Recent studies have suggested similarities between cancer stem cells and the epithelial mesenchymal transition (EMT) state. Our studies suggest that both normal and malignant breast stem cells exist in distinct, inter-convertible states (EMT and MET), the inter-conversion of which is regulated by microRNAs. EMT-like CSCs have a mesenchymal morphology, are largely quiescent, invasive and characterized by expression of the CSC markers CD24^-CD44⁺ and are EpCAM^-CD49f⁺. In contrast, the MET (mesenchymal epithelial transition) state of CSCs is characterized by active self-renewal and expression of the CSC markers ALDH and EpCAM⁺CD49f⁺. A subpopulation of cells expressing both CD24^-CD44⁺ and ALDH may represent cells in transition between these states. This transition is regulated by signals originating in the microenvironment which in turn modulate microRNA networks in the CSC populations. The existence of multiple stem cell states suggests the necessity of developing therapeutic strategies capable of effectively targeting CSCs in all of these states. In addition, since CSC states are regulated by miRNAs, these small non-coding RNAs may be useful therapeutic agents to target CSCs.     

Biological and Genetic Characteristics of Tumor-Initiating Cells in Colon Cancer

Background Human prominin-1 (PROM1, CD133) was used as a marker to detect stem cells (progenitor cells) and cancer stem cells (tumor-initiating cells) in various tissues. The purpose of this study was to investigate the biological and genetic characteristics of tumor-initiating cells in colon cancer with both in vitro and in vivo analyses. Methods The CD133 expression of 12 colon cancer cell lines was evaluated. CD133⁺ cells were isolated by flow cytometry and examined for in vivo tumor formation, in vitro proliferation, colony formation, and invasion ability. Additionally, we used microarray analysis to compare gene expression profiles between CD133⁺ and CD133^– isolated cells. Results CD133⁺ cells were found in 5 of 12 colon cancer cell lines. Isolated CD133⁺ cells from the HT29 colon cancer cell line exhibited a higher tumorigenic potential than CD133^– cells in the in vivo tumor formation assay. Furthermore, it was shown that CD133⁺ cells are more proliferative and have higher colony-forming and invasive abilities than CD133^– cells in vitro. Microarray analysis found differential gene expression correlating with CD133 expression. Conclusions It was confirmed that CD133⁺ cells in colon cancer are useful markers for the detection of tumor-initiating cells. Intimate biological and genetic features of CD133⁺ cells in colon cancer cell lines were also revealed. The biological characteristics of CD133⁺ cells and differentially expressed genes in these cells will help elucidate more details of tumor-initiating cells in colon cancer. Electronic supplementary material The online version of this article (doi: ) contains supplementary material, which is available to authorized users     

Human prostate cancer stem cells: new features unveiled

Cancer stem cells (CSCs) are a rare sub-population of phenotypically distinct cancer cells exhibiting stem cell characteristics.They are tumourigenic, meanwhil...     

Luminal A型和HER-2阳性乳腺癌的肿瘤干细胞生物学行为比较

目的:比较人Luminal A型和HER-2阳性乳腺癌的肿瘤干细胞的生物学行为差异。方法:用Luminal A型和HER-2阳性乳腺癌组织培分离并培养原代乳腺癌细胞,获取富含肿瘤干细胞的微球体(MS);用胰酶将MS消化成单细胞(微球体细胞,MSDC),比较两类乳腺癌MSDC的体外增殖和自我更新能力、ALDH1+表型肿瘤干细胞含量,以及移植NOD/SCID小鼠后的成瘤情况。结果:Luminal A型和HER-2阳性乳腺癌组织分离的原代乳腺癌细胞均能培养出MS,但相对于Luminal A型乳腺癌,HER-2阳性乳腺癌来源的MS形成时间短,生成的MS体积大;HER-2阳性乳腺癌来源的MSDC无论在无血清或添加血清的培养基中的增殖与再生能力均明显强于Luminal A型乳腺癌来源的MSDC,且前者含ALDH1表型干细胞比例明显高于后者(均P0.05);HER-2阳性乳腺癌的MSDC在NOD/SCID小鼠体内成瘤能力也强于Luminal A型乳腺癌来源的MSDC。结论:Luminal A型和HER-2阳性乳腺癌分离出来肿瘤干细胞生物学行为存在明显的差异,两者可能有不同肿瘤干细胞起源。     

CD133作为肝癌干细胞表面标记的初步研究

目的 研究人肝癌SMMC7721和bcl-7402细胞株中CD133的表达,探讨CD133作为肝癌干细胞表面标记的可能性.方法 采用流式细胞仪检测SMMC7721和bcl-7402细胞株中CD133的表达;免疫磁珠细胞技术分离SMMC7721和bcl-7402细胞株中CD133阳性和CD 133阴性细胞,检测其细胞周期,观察并采用单因素方差分析和u检验分析体外培养过程中CD133阳性和CD133阴性细胞的细胞形态、增殖和分化能力的差异.结果 CD133阳性细胞在SMMC7721和bcl-7402细胞株中所占比例分别为0.7%～1.0%、1.7%～8.9%;流式细胞仪检测显示两种细胞株中处于G0/G1期的CD133阳性细胞分别为85.3%、89.4%,明显高于CD133阴性细胞和未分选细胞(F=14.49,38.84,P<0.05);CD133阳性细胞体外增殖能力强于相同条件下CD133阴性细胞和未分选细胞,在1～3 d和5～7 d曲线变化更明显(F=49.32,784.04,89.91,152.83,P<0.05);体外培养过程中,CD133阳性细胞的比例逐日下降,至扩增的第15天,与未分选细胞含量相似(u=0.271,P<0.05).结论 人肝癌SMMC7721和bcl-7402细胞株中,CD133阳性细胞体外增殖和分化能力较强,CD133是肝癌干细胞的表面标记之一.     

胰腺癌肿瘤干细胞对抗肿瘤药物敏感性的实验研究

目的 探讨胰腺癌肿瘤干细胞对抗肿瘤药物的敏感性.方法 FACS技术分选人胰腺癌PANC-1细胞;RT-PCR技术检测分选PANC-1细胞中CD133、ABCG2、Notch1的表达情况;MTT法检测分选细胞对抗肿瘤药物5-氟尿嘧啶和吉西他滨的耐药性.建立分选细胞的移植瘤模型,随机分为吉西他滨治疗组(n=3)和对照组(n=3),观察肿瘤生长情况,作CD133免疫组化染色.结果PANC-1细胞中含有SP亚群.SP细胞CD133、ABCG2、Notch1的mRNA的表达明显上调,non-SP细胞的表达量显著低于前者.在吉西他滨的干预下,SP细胞和non-SP细胞的OD值差异有统计学意义.而5-氟尿嘧啶的干预(10 μg/ml和100 μg/ml)则没有显著差异.移植肿瘤治疗组中CD133阳性细胞明显多于对照组(P=0.001).结论胰腺癌中存在SP亚群细胞.胰腺癌PANC-1的成瘤能力是由其中的SP亚群细胞决定的,而并非所有胰腺癌PANC-1细胞.胰腺癌肿瘤干细胞对抗肿瘤药吉西他滨具有较高耐药性.     

树突状细胞瘤苗联合吉西他滨对肝癌干细胞的杀伤效应

目的:探讨负载CD133+肝癌细胞抗原的树突状细胞(DC)联合吉西他滨(GEM)在体外对肝癌干细胞的杀伤效应。方法:取对数生长期的人肝癌细胞系Huh-7以CD133作为分子标志进行流式分选,得到肝癌干细胞。将人外周血单个核细胞(PBMC)在体外诱导分化为树突状细胞(DC)。DC负载Huh-7细胞和CD133+细胞抗原后与T细胞共育得到特异性细胞毒性T细胞,将CD133+细胞作为靶细胞进行细胞毒性试验。实验组按处理因素分为:GEM组,CD133+-CTL组,GEM+CD133+-CTL组,Huh7-CTL组和GEM+Huh7-CTL组。CCK-8法检测杀伤率,然后比较各组间差异。结果:对CD133+细胞的杀伤效应以GEM+CD133+-CTL组最强,与其他各组比较差异均有统计学意义(P0.05)。单独就DC瘤苗杀伤率,CD133+-CTL组高于Huh7-CTL组(P0.05)。结论:CD133+肝癌干细胞裂解产物致敏的DC瘤苗联合化疗药物可以有效杀伤肝癌干细胞,进而可能降低肝癌术后和肝癌肝移植后的转移和复发率。     

PIWIL2在人膀胱移行细胞癌肿瘤干细胞样细胞中的表达及意义

目的 研究PIWIL2在人膀胱癌肿瘤干细胞样细胞(CSLC)中的表达和意义,探讨PIWIL2在靶向肿瘤干细胞治疗中的作用.方法 用无血清悬浮培养法从人膀胱移行细胞癌细胞株BIU-87中分离获得悬浮细胞球,流式细胞仪检测细胞表面分子标志CD133和CD44的表达,免疫磁珠分选系统分离CD133+CD44+细胞;分别采用T...     

Effective enrichment of prostate cancer stem cells from spheres in a suspension culture system

BackgroundStem-like prostate cancer cells are also called prostate cancer stem cells (PrCSCs). These rare cells are supposed to be highly tumorigenic and to be involved in maintenance of tumor homeostasis and mediation of tumor metastasis. Methods for sorting PrCSCs are mainly based on sorting cells with the marker (CD133⁺/CD44⁺) or side population cells. However, CD133⁺/CD44⁺ cells or side population cells are very rare or even undetectable. The scarcity of approaches for isolation and purification of PrCSCs is the main obstacle to studying PrCSCs.MethodsIn the present study, suspension culture was used for enrichment of PrCSCs. And PrCSCs were verified by side population technology, drug sensitivity assays, and the molecular marker analysis of prostate cancer stem cell.ResultsPC3 cells survived and formed spheres in nonadherent suspension culture. The percentage of CD44⁺/CD133⁺ cells was 18-fold higher in the nonadherent sphere-forming cell population than in the adherent PC3 cell population (13.94% vs. 0.77%, respectively). This side population was increased to 3.1% in the nonadherent population but undetectable in adherent population. Resistance to cisplatin was higher in the nonadherent cells than adherent cells.ConclusionSuspension culture can be used to enrich for PrCSCs. This approach will aid prostate stem cell biology research and facilitate identification of novel therapeutic agents for prostate cancer.     

Single‐marker identification of head and neck squamous cell carcinoma cancer stem cells with aldehyde dehydrogenase

Background

In accord with the cancer stem cell (CSC) theory, only a small subset of cancer cells are capable of forming tumors. We previously reported that CD44 isolates tumorigenic cells from head and neck squamous cell cancer (HNSCC). Recent studies indicate that aldehyde dehydrogenase (ALDH) activity may represent a more specific marker of CSCs.

Methods

Six primary HNSCCs were collected. Cells with high and low ALDH activity (ALDH^high/ALDH^low) were isolated. ALDH^high and ALDH^low populations were implanted into NOD/SCID mice and monitored for tumor development.

Results

ALDH^high cells represented a small percentage of the tumor cells (1% to 7.8%). ALDH^high cells formed tumors from as few as 500 cells in 24/45 implantations, whereas only 3/37 implantations of ALDH^low cells formed tumors.

Conclusions

ALDH^high cells comprise a subpopulation cells in HNSCCs that are tumorigenic and capable of producing tumors at very low numbers. This finding indicates that ALDH activity on its own is a highly selective marker for CSCs in HNSCC. © 2010 Wiley Periodicals, Inc. Head Neck, 2010     

Chemoresistance to 5-FU inhibited by 635 nm LED irradiation in CD133+ KB cell line

Consistent with cancer stem cell theory, a small fraction of cancer cells, described as cancer stem cells (CSCs), may promote tumor recurrence and anti-cancer drug resistance. Therefore, much effort has been devoted to the development of CSC targeted therapy to vanquish drug resistance. In this study, we have investigated the effect of multiple light-emitting diode (LED) irradiation treatments with conventional anti-cancer drugs on CSC-like oral cancer cells that acquired stemness by ectopic over expression of CD133. To evaluate combined LED irradiation anti-cancer drug effects, we investigated the chemosensitizing effect of 635 nm irradiation on 5-fluorouracil (5FU)-treated KB^CD133+ and KB^Vec cells, interrogating the underlying molecular mechanisms associated with stemness and apoptosis that are responsible for chemopreventive activity. In addition, combination therapy with LED irradiation and 5-FU treatment was carried out in KB^CD133+ and KB^Vec cell-inoculated mouse models. LED irradiation of 635 nm inhibited CSC-like properties consistent with a decrease in OCT4 and NANOG protein expression, reducing colony-forming ability. In addition, LED irradiation enhanced 5-FU-induced cytotoxicity and improved 5-FU chemosensitivity in KB^CD133+ via enhancement of apoptosis. These findings were validated in vivo, wherein LED irradiation combined with 5-FU treatment inhibited tumor growth in KB^CD133+-inoculated mice. Collectively, our results provide novel evidence for 635 nm irradiation-induced 5-FU chemosensitization of CSC in oral cancer. In addition, this research highlights that 635 nm LED irradiation may serve as an adjunct treatment to conventional chemotherapeutic drugs in patients with oral cancer.     

CD44+-ESA+在结肠癌干细胞分选中的应用

目的 分选结肠癌细胞株SW480细胞中的CD133+-CD44+-ESA+亚群细胞,并观察其致瘤性.方法 用流式细胞仪分选SW480细胞中CD133+-CD44+-ESA+、CD133--CD44+-ESA+及CD133--CD44--ESA-亚群细胞.将这3组细胞分别接种于NOD/SCID小鼠,每组5只,观察肿瘤生长...     

The role of cancer stem cells in immunotherapy for bladder cancer: An in vitro study

ObjectiveBladder cancer is characterized by frequent recurrence and progression. CD44+ cancer stem cells (CSCs) might be one of the main reasons for recurrence. Although Bacillus Calmette Guerin (BCG) has become a gold standard immunotherapy, after treatment recurrence frequently occur. Based on this knowledge, the aim of this study was to evaluate the changes in cytokine and chemokine expressions in bladder cancer and CSCs cultures in vitro with BCG only and in combination with IL2 and lymphocyte (MNCs) applications.Material and methodsIn this study, 3 cell lines of human bladder cancer cells with different characteristics (T24, 5637, and JMSU-1) and CD44+ bladder CSCs isolated by magnetic bead isolation (Miltenyl Magtech) were used. Bladder cancer cell lines and bladder CSCs in complete medium were cultured under humidified conditions of 37°C temperature in 5% CO₂. BCG only and its combination with IL2 and MNCs were applied to bladder cancer cell lines and bladder CSCs for 24, 48, and 72 hours. Annexin V-PI was used to detect the percentages of apoptotic and necrotic cells in treatment groups and control groups. After treatments, total RNAs were isolated and converted to cDNA for each group and controls. Quantitative fold changes in terms of gene expression were measured by RT2–PCR array and fold changes for expression levels of genes were compared among groups. Eighty-four genes were analyzed in standard array of chemokines and cytokines (Biorad).ResultsBCG treatment with 7.32 µg/ml dose alone and in combination with IL2 (1000 IU/ml) and MNCs (1000 cells/ml) were found to be most effective on bladder cancer cells. When BCG and its combinations were applied to CSCs of the 3 cell lines, BCG treatment showed cytotoxic effect on CSCs as well as cancer cells. CSCs of 3 cell lines over expressed CXCL5, CCL8, CNTF, and CSF2 compared with cancer cells. Cancer cells over expressed IL6, TNSFF11, FASLG, and CXCL9 compared with CSCs. In all 3 cell lines, BCG application increased expression of CXCL5 and LTB and also decreased CCL20 and IL6. When BCG was combined with IL2 and MNCs, CXCL10, CXCL5, and IFNG were increased and CXCL12, IL6, and TNSF11 were decreased. BCG treatment of CSCs caused increases in ADIPOQ, CXCL10, and XCL1 and a decrease in CCL8. When IL2 and MNCs were combined with BCG, the expression of many cytokines and chemokines decreased.ConclusionBCG treatment changes the expression of many cytokines and chemokines in bladder cancer. The expression differs in 3 different cell lines and their CSCs. Immune modulation of each case differs from each other. The effectivity of BCG-based immunotherapy in bladder cancer on CSCs might decrease in combination with IL2. Our results indicate that recurrence after BCG treatment for bladder cancer may not occur mainly based on the CSCs hypothesis considering bladder cancer occurs at different loci of surface epithelium.     

CD133<Superscript>+</Superscript>CD44<Superscript>+</Superscript> Population Efficiently Enriches Colon Cancer Initiating Cells

Background  Previous reports have demonstrated that CD133⁺ cells or CD44⁺ cells might be cancer initiating cells (CIC) of colon cancer. However, the association between the two cell types is unclear. In this study, we evaluated the tumorigenicity of each population of human colon cancer divided by CD133 and CD44 using non-obese diabetic/severe combined immunodeficient (NOD/SCID) mice. Methods  Using the colon cancer cell lines HT29 and Caco2 we evaluated the change of expression status of CD133 or CD44 by a treatment with sodium butyrate (NaBT) that can induce cellular differentiation. Next, we prepared ten clinical samples of colon cancer and analyzed the expression and tumorigenicity of CD133 and CD44. Results  With NaBT treatment, CD44 expression was greatly downregulated in both HT29 and Caco2 (HT29: nontreatment versus treatment; 77.8% versus 0.6%, Caco2: 14.0% versus 0.4%, respectively), more than CD133 expression (HT29: nontreatment versus treatment; 90.1% versus 67.7%, Caco2: 98.9% versus 76.3%, respectively). In clinical samples, the percentages of CD133⁺ cells and CD44⁺ cells varied from 0.3% to 82.0% (mean 35.5%), and from 11.5% to 58.4% (mean 30.0%), respectively. Subcutaneous injection of CD133⁺ or CD44⁺ cells made a tumor in all mice (3/3 and 4/4, respectively). The combined analysis of CD133 and CD44 revealed that only the CD133⁺CD44⁺ population had the ability to produce a tumor (3/3). Conclusion  The findings demonstrate that, at present, the CD133⁺CD44⁺ population may be the best to identify tumor initiating cells of human colon cancer. Naotsugu Haraguchi: Awardee of Research Resident Fellowship from the Foundation for Promotion of Cancer Research (Japan) for the 3^rd Term Comprehensive 10- Year Strategy for Cancer Control.     

The role of cancer stem cells (CD133(+)) in malignant gliomas

Malignant gliomas, particularly glioblastoma multiforme (GBM) tumors, are very difficult to treat by conventional approaches. Although most of the tumor mass can be removed by surgical resection, radiotherapy, and chemotherapy, it eventually recurs. There is growing evidence that cancer stem cells (CSCs) play an important role in tumor recurrence. These stem cells are radioresistant and chemoresistant. The most commonly used tumor marker for CSCs is CD133. The amount of CSC component is closely correlated with tumor malignancy grading. Isolating, identifying, and treating CSCs as the target is crucial for treating malignant gliomas. CSC-associated vascular endothelial growth factor (VEGF) promotes tumor angiogenesis, tumor hemorrhage, and tumor infiltration. Micro-RNA (miRNA) plays a role in CSC gene expression, which may regulate oncogenesis or suppression to influence tumor development or progression. The antigenesis of CSCs and normal stem cells may be different. The CSCs may escape the T-cell immune response. Identifying a new specific antigen from CSCs for vaccine treatment is a key point for immunotherapy. On the other hand, augmented treatment with radiosensitizer or chemosensitizer may lead to reduction of CSCs and lead to CSCs being vulnerable to radiotherapy and chemotherapy. The control of signaling pathway and cell differentiation to CSC growth is another new hope for treatment of malignant gliomas. Although the many physiological behavioral differences between CSCs and normal stem cells are unclear, the more we know about these differences the better we will be able to treat CSCs effectively.