CXCR 4CD44 CD133表达在舌鳞状细胞癌患者生存分析中的价值

目的:探讨影响舌鳞状细胞癌患者术后生存的相关因素。方法:回顾性分析经病理确诊并行手术治疗的44例舌鳞癌患者临床资料,采用免疫组织化学方法检测不同病理分级舌鳞癌患者癌组织中CXCR4、CD44、CD133的表达情况。将可能影响患者术后生存的指标进行Kaplan-Meier检验后,采用Cox比例风险回归模型进行多因素分析。结果:本研究44例舌鳞癌标本中,高分化29例,中、低分化15例;Ⅰ期11例,Ⅱ期12例,Ⅲ期8例,Ⅳ期13例。各病理分级病例CXCR4、CD44、CD133阳阳性率分别是79.54%(35/44)、77.27%(34/44)和75.00%(33/44)。CXCR4、CD44、CD133在舌鳞癌各病理分级组在之间表达强度差异均有统计学意义(P<0.05),且CXCR4、CD44、CD133分别与转移、复发也成正相关。Cox模型多因素分析提示:CXCR4表达情况、临床分期、颈部转移为本组舌鳞癌患者预后独立的影响因素及死亡的危险因素。结论:CXCR4、CD44、CD133的表达与舌鳞癌的恶性程度存在相关性,CXCR4表达情况、临床分期、颈部转移为术后评价舌鳞癌患者生存的重要指标。

The human leucocyte differentiation antigens (HLDA) workshops: the evolv-ing role of antibodies in research, diagnosis and therapy

The 8^th International Workshop on Human Leucocyte Differentiation Antigens （chaired by HZ and managed by BS） was run over a 4-year period and culminated in a conference in December 2004. Here we review the achievements of the HLDA Workshops and provide links to information on CD molecules and antibodies against them, including the 93 new CDs assigned in the 8^th Workshop. We consider what remains to be achieved （including an estimate of the number of leucocyte surface molecules still to be discovered）, and how the field can best move forward.

The Cross-talk between ROS and p38MAPKα in the Ex Vivo Expanded Human Umbilical Cord Blood CD133^+ Cells

This study investigated the correlation between and compared the effects of reactive oxygen species（ROS） and p38 mitogen-activated protein kinase α（p38MAPKα） in the ex vivo expanded umbilical cord blood（hUCB） CD133＋ cells.hUCB CD133＋ cells were cultured in the hematopoietic stem cells（HSCs） culture medium with N-acetylcysteine（NAC,an anti-oxidant）,p38MAPKα-specific inhibitor（SB203580） or their combination.The levels of ROS and expression of phosphorylated p38MAPKα（p-p38） in CD133＋ cells were flow cytometrically detected.The efficacy of ex vivo expansion was evaluated by the density of CD133＋ cell sub-group colony-forming cells（CFC） and cobblestone area-forming cells（CAFC） assay.Our results showed decreased ROS levels in NAC,SB203580,and their combination treatment groups were almost 37%,48%,and 85%,respectively.Furthermore,SB203580 abrogated the activation of p38MAPKα more obviously than NAC.Moreover,the CD133＋ cells in SB203580 treatment group had a 21.93±1.36-fold increase,and 14.50±1.19-fold increase in NAC treatment group,but only 10.13±0.57-fold increase in control group.In addition,SB203580 treatment led a higher level increase in the number of CFU and CAFC than NAC did.These findings suggested that,in expanded CD133＋ cells,ROS activates p38MAPKα,which,in turn,induces ROS production,and p38MAPKα might be the most suitable regulator in ROS-p38MAPKα pathway for the promotion of HSCs ex vivo expansion.

Autologous CD34^+ and CD133^+ stem cells transplantation in patients with end stage liver disease

AIM:To assess the utility of an autologous CD34 + and CD133 + stem cells infusion as a possible therapeutic modality in patients with end-stage liver diseases.METHODS:One hundred and forty patients with endstage liver diseases were randomized into two groups.Group 1,comprising 90 patients,received granulocyte colony stimulating factor for five days followed by autologous CD34 + and CD133 + stem cell infusion in the portal vein.Group 2,comprising 50 patients,received regular liver treatment only and served as a control group.RESULTS:Near normalization of liver enzymes and improvement in synthetic function were observed in 54.5% of the group 1 patients;13.6% of the patients showed stable states in the infused group.None of the patients in the control group showed improvement.No adverse effects were noted.CONCLUSION:Our data showed that a CD34 + and CD133 + stem cells infusion can be used as supportive treatment for end-stage liver disease with satisfactory tolerability.

Cancer stem cell markers CD133 and CD24 correlate with invasiveness and differentiation in colorectal adenocarcinoma

AIM:To verify that CD markers are available for detecting cancer stem cell populations and to evaluate their clinical significance in colon cancer.METHODS:Immunohistochemistry for CD133,CD24 and CD44 was performed on the tissue microarray of 523 colorectal adenocarcinomas.Medical records were reviewed and clinicopathological analysis was performed.RESULTS:In colorectal adenocarcinoma,128 of 523 cases(24.5%) were positive and 395 cases(75.5%) were negative for CD133 expression.Two hundred and sixty-four of 523 cases(50.5%) were positive and 259 cases(49.5%) were negative for CD24 expression.Five hundred and two of 523 cases(96%) were negative and 21 cases(4%) were positive for CD44 expression.Upon clinicopathological analysis,CD133 expression was present more in male patients(P = 0.002) and in advanced T stage cancer(P = 0.024).Correlation between CD24 expression and clinicopathological factors was seen in the degree of differentiation(P = 0.006).Correlation between CD44 expression and clinicopathological factors was seen in the tumor size(P = 0.001).Survival was not significantly related to CD133,CD24 and CD44 expression.CONCLUSION:CD markers were related to invasiveness and differentiation of colorectal adenocarcinoma.However,CD expression was not closely related to survival.

Upregulated CD133 expression in tumorigenesis of colon cancer cells

AIM: To analyze the upregulated CD133 expression in tumorigenesis of primary colon cancer cells. METHODS: Upregulated CD133 expression in tumorigenesis of colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) was analyzed by flow cytometry. Human colon cancer tissue samples were stained with anti-human CD133. SW620 cells were sorted according to the CD133 expression level measured by fluorescence-activated cell sorting. Spheroids of colorectal cancer cells were cultured with the hanging drop. Expression of CD133 and Lgr5 in spheroids of colorectal cancer cells and monolayer culture was detected by RT-qPCR. Spheroids of colorectal cancer cells were analyzed using anti-human CD133 with immunohistochemical staining. RESULTS: CD133 antigen was expressed in colorectal cancer cell lines (Lovo, Colo205, Caco-2, HCT116 and SW620) as well as in primary and metastatic human colon cancer tissues. However, the CD133 was differently expressed in these cell lines and tissues. The expression levels of CD133 and Lgr5 were significantly higher in spheroids of parental, CD133hi and CD133-cells than in their monolayer culture at the mRNA level (P < 0.05). Immunohistochemical staining of spheroids of CD133-cells showed that CD133 was highly expressed in colorectal cancer cell lines. CONCLUSION: Upregulated CD133 expression plays a role in tumorigenesis colorectal cancer cells, which may promote the expression of other critical genes that can drive tumorigenesis.

Promoter hypomethylation regulates CD133 expression in human gliomas

Brain tumor-initiating cells （BTICs） have been enriched using antibodies against the cell surface protein CD133; however, the biological relevance and the regulatory mechanism of CD133 expression in human gliomas are not yet understood. In this study, we initially demonstrated that CD133 was overexpressed in high-grade human glioblastomas where CD133-positive cells were focally observed as a micro-cluster. In addition, CD133 transcripts with exon 1A, 1B, or 1C were predominantly expressed in glioblastomas. To elucidate the mechanism regulating this aberrant expression of CD133, three proximal promoters （P1, P2, and P3） containing a CpG island were isolated. In U251MG and T98G glioblastoma cells, the P1 region flanking exon 1A exhibited the highest activity among the three promoters, and this activity was significantly inactivated by in vitro methylation. After treatment with the demethylating agent 5-azacytidine and/or the histone deacetylase inhibitor valproic acid, the expression level of CD133 mRNA was significantly restored in glioma cells. Importantly, hypomethylation of CpG sites within the P1, P2, and P3 regions was observed by bisulfite sequencing in human glioblastoma tissues with abundant CD133 mRNA. Taken together, our results indicate that DNA hypomethylation is an important determinant of CD133 expression in glioblastomas, and this epigenetic event may be associated with the development of BTICs expressing CD133.

Promoter hypermethylation and loss of CD133 gene expression in colorectal cancers

AIM: To understand CD133 promoter hypermethyl-ation and expression in 32 colorectal cancer cell lines. METHODS: Nucleic acid was isolated from 32 colorectal cancer cell lines and CD133 expression levels were measured by reverse transcription-polymerase chain reaction (RT-PCR) and real-time PCR. Promoter methylation status of the CD133 gene was analyzed with a methylation-specific PCR after sodium-bisulfi te modification and by clonal sequencing analysis. The correlation between expression and promoter methylation of CD133 gene was confirmed with treatment of 5-aza-2’-deoxycytidine. RESULTS: We measured CD133 expression levels in 32 colorectal cancer cell lines. RT-PCR analysis showed undetectable or low levels of CD133 expression in 34.4%of cell lines. To verify the relation between CD133 expression and methylation status of the CD133 gene promoter in colorectal carcinogenesis, CD133 gene promoter hypermethylation was analyzed in 32 cancer cell lines. Promoter hypermethylation was detected in 13 (40.6%) of the cell lines using methylation specificPCR and confirmed by bisulfite sequencing analysis. Treatment of 11 of the cell lines with the demethylation agent 5-aza-2’-deoxycytidine recovered CD133 expression in most of them. CONCLUSION: Transcriptional repression of CD133 is caused by promoter hypermethylation of the CD133 CpG islands in some of colorectal cancer cell lines. The study may contribute to the understanding of the role of CD133 inactivation in the progression of colorectal cancers.

Strong expression of CD133 is associated with increased cholangiocarcinoma progression

AIM:To determine the role of CD133 in cholangiocarcinoma progression. METHODS:CD133 protein expression was evaluated by immunohistochemistry in 34 cholangiocarcinoma specimens.In addition,proliferation,chemoresistance and invasive properties of CD133-enriched(CD133 + ) and CD133-depleted(CD133 )RMCCA1 cholangiocarcinoma cells were studied and compared. RESULTS:Strong CD133 expression was observed in 67.6%(23/34)of the cholangiocarcinoma specimens. Strong expression of CD133 was significantly associated with nodal metastasis(P=0.009)and positive surgical margin status(P=0.011).In the in vitro study, both the CD133 + and CD133 cells had similar proliferation abilities and resistance to chemotherapeutic drugs.However,the CD133 + cells had a higher invasive ability compared with CD133 cells. CONCLUSION:CD133+cells play an important role in the invasiveness of cholangiocarcinoma.Targeting of the CD133+cells may be a useful approach to improve treatment against cholangiocarcinoma.

Inhibitive effect of IL-24 gene on CD133^+ laryngeal cancer cells

Objective:To explore the inhihitive and apoptosis inductive effect of IL-24 genes on CD133^+laryngeal cancer cells in Hep-2 line.Methods:Human peripheral blood monocytes were isolated.The total RNA was extracted by using Trizol method and reverse transcripted into cDNA using RT-PCR method.Primers P1 and P2 was designed for the amplification of human IL-24 genes.After confirmation of agarose gel electrophoresis tests,TA was cloned into pMD19-T simple vector.Nhe Ⅰ and Xho Ⅰ double digesting human IL-24 and pIRES2-ZsGreen1 and eukaryotic expression vector were used to establish the pIRES2-ZsGreen1-hIL-24 vector,and detected by enzyme digestion and gene sequencing methods.Flow cytometry(FCM) was used to isolate CD133^+ cells from Hep-2 cells.CD133^+ cells were transfected with pIRES2-ZsGreen1-hIL-24 through liposome 2000.After detection,MTT and FCM were used to observe the effect of IL-24 gene on CD133^+ laryngeal cancer Hep-2 cells.Results:Lipotin mediated transfection of recombinant pIRES2-ZsGreen1-hIL-24 plasmid into CD133^+ Hep-2 could expressed IL-24 gene in cells stably.MTT results showed that IL-24 transfected group was significantly suppressed compared to empty vector group and control group(P<0.05);FCM results showed that the apoptosis rate of experimental group increased significantly compared to empty vector group and control group(P<0.05).Conclusions:IL-24 gene expressions can inhibit proliferation of CD133^+laryngeal cells in Hep-2 line and promote their apoptosis.

Angioarchitecture and CD133^+ tumor stem cell distribution in intracranial hemangiopericytoma A comparative study with meningioma

Angioarchitecture plays an important role in the malignant development of intracranial hemangiopericytoma. It remains poorly understood whether high frequency of hemorrhage during clinical surgery for intracranial hemangiopericytoma is associated with angioarchitecture. The present study utilized hematoxylin-eosin staining, and immunohistochemical staining with epithelial membrane antigen, vimentin, CD34, von Willebrand factor （vWF） and CD133 to observe characteristics of angioarchitecture. In addition, silver stains were used to demonstrate changes in reticular fibers in the wall of vessel channels in intracranial hemangiopericytoma and meningioma. Five patterns of angioarchitecture were identified in intracranial hemangiopericytoma, namely tumor cell islands, vasculogenic mimicry, mosaic blood vessels, sprouting angiogenesis, and intussusceptive angiogenesis. Several CD133＋ tumor cells were found to form tumor cell islands. A connection between vWF ^＋ and vWF channels was detected in the pattern of intussusceptive angiogenesis, and some vimentin^＋ tumor cells were embedded in the periodic acid-Schiff positive channel wall. Incomplete threads of reticular fibers formed the walls of larger pseudo-vascular channels and some tumor clumps or scattered tumor cells were detected ＂floating＂ in them. The angioarchitecture, specific markers and reticular fibers of intracranial hemangiopericytoma were significantly different from meningioma. Angioarchitecture provides a functional vascular network for vascular evolution in intracranial hemangiopericytoma and contributes to significant intra-operative bleeding.

Cisplatin selects for CD133+cells in lung cancer cells

Objective Platinum-based chemotherapy is the first-line treatment for non-small cell lung cancer,but the chemoresistance of tumor cells continues to be a considerable challenge in the management of NSCLCs,leading to recurrence of most patients.CD133(prominin-1)is a five-transmembrane glycoprotein,and recent evidence suggests that CD133+cells are the cause of drug resistance and tumor recurrence.In this study,the correlation between cisplatin and CD133+cells was investigated systematically.Methods Four lung cancer cell lines,including A549,H460,801D and H1299,were treated with different concentrations of cisplatin.Cell viability was determined by MTT assay.Sphere-forming assay was performed to detect the capability of sphere-forming.CD133+cells was detected by BD FACScaliber flow cytometer.Results The results showed that cisplatin could increase the number of CD133+cells in both time-and dose-dependent manner.The enrichment would weaken but the proportion of CD133+cells was still higher than the basic level as incubation time extended after cisplatin was withdrawn.Compared with adherent culture,the proportion of CD133+cells was higher when the cells were maintained suspension culture.The proportion of CD133+cells significantly increased when cisplatin was provided in suspension culture.Conclusion These results revealed that cisplatin induces the enrichment of CD133+cells and CD133 is a new therapeutic target.Our data partially explained drug resistance to second-line chemotherapy in cisplatin-treated patients with NSCLCs.

Influence of CD133^+ expression on patients' survival and resistance of CD133^+ cells to anti-tumor reagents in gastric cancer

Objective:To investigate the influence of CD133+expression on patients'survival and resistance of CD133+cells to anti-tumor agents in gastric cancer(GC).Methods:Influence of CD133 expression on prognosis was analyzed employing samples from patients with GC.GC cell lines were utilized to separate CD133+and CD133-subpopulations by immunomagnetic separation and to analyze the biological features of two subpopulations in vitro and in vivo,especially in resistant to anti-tumor reagents and its apoptotic mechanism.Results:The lower CD133+group showed a significantly better survival compared with the higher CD133+group.The highest content of CD133+subpopulations for KATO-III cells had stronger proliferative ability than CD133-subpopulations.A single CD133+cell was capable of generating new cell colony and the tumorigenicity rate in nude mice was100%for CD133+clonal spheres or for CD133+cells,but 0%for CD133-cells.Furthermore,the higher expression levels of Oct-4,Sox-2,Musashi-1 and ABCG2 in CD133+clonal spheres were identified compared with CD133+cells or CD133-cells.Under the treatment of anti-tumor reagents,CD133+cells had lower suppression rates compared with CD133-cells while lower level of Bcl-2 and higher level of Bax were found in CD133+cells compared with CD133-cells.Conclusions:The patients with lower CD133+expression had a better survival.Enriched CD133+cells in clonal sphere shared the ability to be self-renewable,proliferative,tumorigenic and resistant to anti-tumor agents as probably regulated by Bcl-2 and Bax.

Oxidized low-density lipoprotein stimulates CD206 positive macrophages upregulating CD44 and CD133 expression in colorectal cancer with high-fat diet

BACKGROUND Oxidized low-density lipoprotein(ox-LDL),which is abnormally increased in the serum of colorectal cancer(CRC)patients consuming a high-fat diet(HFD),may be one of the risk factors for the development of CRC.Ox-LDL exerts a regulatory effect on macrophages and may influence CRC through the tumor microenvironment.The role of ox-LDL in CRC remains unclear.AIM To investigate the role of ox-LDL through macrophages in HFD associated CRC.METHODS The expression of ox-LDL and CD206 was detected in colorectal tissues of CRC patients with hyperlipidemia and HFD-fed mice by immunofluorescence.We stimulated the macrophages with 20μg/mL ox-LDL and assessed the expression levels of CD206 and the cytokines by cell fluorescence and quantitative polymerase chain reaction.We further knocked down LOX-1,the surface receptor of ox-LDL,to confirm the function of ox-LDL in macrophages.Then,LoVo cells were co-cultured with the stimulated macrophages to analyze the CD44 and CD133 expression by western blot.RESULTS The expression of ox-LDL and the CD206 was significantly increased in the stroma of colorectal tissues of CRC patients with hyperlipidemia,and also upregulated in the HFD-fed mice.Moreover,an increased level of CD206 and decreased level of inducible nitric oxide synthase were observed in macrophages after ox-LDL continuous stimulation.Such effects were inhibited when the surface receptor LOX-1 was knocked down in macrophages.Ox-LDL could induce CD206+macrophages,which resulted in high expression of CD44 and CD133 in co-cultured LoVo cells.CONCLUSION Ox-LDL stimulates CD206+macrophages to upregulate CD44 and CD133 expression in HFD related CRC.

Proliferation Characteristics of CD133+ Cell Population in Colorectal Cancer

In this study,CD133+ subpopulations were isolated from 41 primary colorectal cancer tissues,the proliferation and cell cycle distribution of the cells were examined without in vitro expansion,and then compared to those of cell lines.The detection of CD133 in colorectal cancer tissues,isolation of CD133+ and CD133-epithelial subpopulations,Ki-67/DNA multiparameter assay and cell volume analysis were flow cytometrically conducted.The results showed that Ki-67 expression was correlated with CD133 level in primary cancer tissues,while cell cycle G 2 /M phase distribution or clinicopathological characteristics was not.In addition,the CD133+ cells showed larger cell volume and higher Ki-67 expression as compared with CD133-cells.But there was no statistically significant difference in G 2 /M phase distribution between the two subpopulations.Our results demonstrated that the CD133+ subpopulation in colorectal cancer tissue contained more actively cycling and proliferating cells,which was not correlated to clinicopathological factors but might contribute to tumor progression and poor clinical outcome.

Colorectal cancers with aneuploids show high CD133 expression and poor prognosis

Objective： The aim of the study was to investigate the relationship of DNA ploidy status in colorectal cancers with patients＇ prognosis and also the relationship of DNA ploidy status with expression of the colorectal cancer stem cell marker CD133. Methods： The DNA ploidy status and CD 133 expression in colorectal cancers were detected by flow cytometry. The clinicopathological characteristics and progression-free survival analysis of patients was evaluated based on the clinical data. Results： DNA ploidy pattern did not correlated with gender, age, lesion diameter, histological type, depth of tumor invasion, lymphatic invasion and Dukes stage. Only primary lesion cite showed significant correlation with DNA ploidy pattern, more aneuploids were observed in colonic cancer than rectal cancer, P 〈 0.05. The 2-year progress/on-free survival rate and total progression-free time in patients with aneuploids were lower than that with diploids, P 〈 0.05. Tumors contained aneuploids showed higher expression of CD133 than tumors of only diploids, P 〈 0.05. Conclusion： Tumor DNA ploidy status is a significant prognostic factor in patients with colorectal cancer and also associated with the existence of CD133 positive colorectal cancer stem cells.

Influence on Biological Behavior of Colon Cancer Stem Cells after RNA Interfering CD133

OBJECTIVE To observe the effects expression and activation on biological cancer stem cells. of blocking CD133 gene characteristic of the colon METHODS CD133＋ colon cancer stem cells （CCSCs） were separated from EpCAMhigh CD44＋ CCSCs through fluorescenceactivated cell sorting （FACS）. The proliferation, the capability of spherical cell formation, neoplasia, and the expression of ABCG2 mRNA of CD133＋ CCSCs were observed after the CD133＋ CCSCs were infected with LV-CD133shRNA. CD133 negative cells were isolated from EpCAMhigh CD44＋ CCSCs with FACS, and the CD133 proteins in CD133- cells were detected with Western blot. RESULTS CD133＋ CCSCs were isolated from EpCAMhigh CD44＋ CCSCs using FACS, and they accounted for 89.2% in the stem cells. In the experimental group, after the CD133＋ CCSCs were knocked down by LV-CD133shRNA RNAi, the growth pattern of the cells in the stem cell culture changed into adherent growth from suspended growth, and couldn＇t generate spherical cells. Results of MTT assay showed that the CD133＋ CCSCs infected with LV-CD133shRNA grew slowly, compared to the cells in the control groups. There was a decrease in the cloning efficiency. The infected cells were transplanted into the BALB/c nude mice. During the observation, no neoplasia was found in the CD133＋ cells infected with LV-CD133shRNA. The level of ABCG2 mRNA expression was lowered greatly （P 〈 0.01）. CD133- cells were obtained from the EpCAMhigh CD44＋ CCSCs using FACS, in which the expression of CD133 protein was positive. CONCLUSION CD133 retains the biological characteristics of the colon cancer stem cells.

Expression of CD133 and Extracellular Matrix Molecules in Malignant Brain Tumors

Background: CD133 could be characterized as a “stem-like” cell subpopulation and an invasive tumor phenotype. The objectives of this study were to investigate the relationship of CD133 and other remodeling factors such as matrix metalloproteinases (MMP) in the brain tumors. Methods: Tumors from 13 patients with brain tumors (8 lung cancer metastasis, 3 breast cancer metastasis, 2 gliomas) were studied to investigate the expression-patterns of CD133, EGFR, MT1-MMP, and MMP7 using the immunostaining and RT-PCR analysis. Results: EGFR immunostaining was detected in 75% (6/8) and 67% (1/3) of brain metastasis from lung adenocarcinoma and breast cancer, respectively. MT1-MMP immunostaining was also detected in 73% (8/11) of these brain metastasis. CD133 was not detected in these 13 patients. EGFR immunostaining was detected in 75% (6/8) and 67% (1/3) of brain metastasis from lung adenocarcinoma and breast cancer, respectively. MT1-MMP immunostaining was also detected in 73% (8/11) of these brain metastasis. CD133 was not detected in these 13 patients. Conclusions: The expression of CD133 indicates a marker for brain tumor initiating.

Prognostic value of cancer stem cell markers CD133, ALDH1 and nuclear β-catenin in colon cancer

Objective： Colon cancer is one of the most common human malignancies. Cancer stem cells （CSCs）, despite being only a small subset of cancer cells, have the capability to self-renew and sustain the tumor. They also have the ability to proliferate. Multiple CSCs-associated markers have been identified in colon cancer including CD133, ALDH1 and β-catenin. The aim of the work was to study the prognostic value of CSCs markers （CD133, ALDH 1 and β-catenin）, as well as their rela- tionship to clinicopathological features of colon cancer. Methods： CD133, ALDH1 and β-catenin proteins expression was as- sessed immunohistochemically in a series of colon cancers and their prognostic significance was evaluated. Results： CD133 expression showed significant relationship to tumor stage and lymph node metastasis （P-value 0.004 ＆ 〈 0.001 respectively）, and near significant relationship to liver metastasis （P-value 0.092）. ALDH1 was significantly associated with tumor grade, stage and nodal metastasis （P-value 0.021,0.001 and 0.026 respectively）, but its relationship to liver metastasis was near sig- nificant （P-value 0.068）. Nuclear β-catenin was significantly related to tumor grade, stage, nodal and liver metastasis （P-value 0.001, 〈 0.001, 〈 0.001 and 0.008 respectively）. Overall survival （OS） was associated inversely with CD133, ALDH1 positivity, and directly with nuclear 13-catenin posiUvity （P-value 〈 0.001,0.0001 and 〈 0.001 respectively）. Also recurrence free survival （RFS） was associated inversely with CD133, ALDH1 and directly with nuclearβ-catenin positivity （P-value 0.0001,0.001 and 〈 0.001 respectively）. Conclusion： CD133, ALDH1 and β-catenin expressions of tumor cells have significant impact upon malignant progression of colon cancer and thus patient survival and tumor recurrence. Hence they can be used to predict outcome of colon cancer patients.