Clinical relevance of stem cell surface markers CD133, CD24, and CD44 in colorectal cancer

Colon cancer stem cells (CSC) identified by cell surface markers CD133, CD24, and CD44, have been shown to be involved with tumor formation, chemotherapy resistance, and the progression of metastatic disease. Using an in silico translational approach, we hypothesize that a combination of these CSC markers has prognostic value in a large cohort of patients with colorectal cancer. Clinicopathologic and RNA expression data from a total of 594 colorectal cancer (CRC) patients from TCGA were analyzed. The expression of CD133, CD24, and CD44 was individually defined as “high” or “low” based on the median expression. Disease specific survival (DSS) and overall survival (OS) were not associated with tumors that are CD133-high or CD44-high alone. Patients with CD24-high tumors have significantly better DSS (P<0.001) and OS (P = 0.043). CD24-high, CD44-high and CD133-high tumors were associated with significantly greater EGFR, KRAS and Ki67 expression (all P<0.001). CD133, CD24 and CD44-high tumors were independently enriched for conventional stemness-related signaling pathways such as Wnt/β-catenin and Hedgehog signaling pathways. There was no survival difference linked to CD133-high/CD44-low patients, but CD44-high/CD24-low patients have worse DSS (P = 0.005) compared with CD44-low/CD24-high patients. CD133-high/CD24-low tumors show significant negative enrichment of MYC targets, E2F targets, G2M checkpoint and mitotic spindle gene sets, suggesting less cell proliferation in these tumors. Patients with CD133-high/CD24-low tumors have worse DSS (P = 0.004) and OS (P = 0.044), and are more likely to have early and late recurrences. In conclusion, we demonstrated that CD133-high/CD24-low tumors may predict colorectal cancer prognosis.     

Prognostic impact of the expression of putative cancer stem cell markers CD133, CD166, CD44s, EpCAM, and ALDH1 in colorectal cancer

Background:

The aim of this study was to elucidate the prognostic impact of putative cancer stem cell markers CD133, CD166, CD44s, EpCAM, and aldehyde dehydrogenase-1 (ALDH1) in colorectal cancer.

Methods:

A tissue microarray of 1420 primary colorectal cancers and 57 normal mucosa samples was immunostained for CD133, CD166, CD44s, EpCAM, and ALDH1 in addition to 101 corresponding whole tissue sections. Invasive potential of three colorectal cancer cell lines was tested.

Results:

Differences between normal tissue and cancer were observed for all markers (P<0.001). Loss of membranous CD166 and CD44s were linked to higher pT (P=0.002, P=0.014), pN (P=0.004, P=0.002), an infiltrating growth pattern (P<0.001, P=0.002), and worse survival (P=0.015, P=0.019) in univariate analysis only. Loss of membranous EpCAM expression was also linked to higher pN (P=0.023) and infiltrating growth pattern (P=0.005). The CD44s, CD166, and EpCAM expression were lost towards the invasive front. The CD44−/CD166− cells from three colorectal cancer cell lines exhibited significantly higher invasive potential in vitro than their positive counterparts.

Conclusions:

Loss, rather than overexpression, of membranous CD44s, CD166, and EpCAM is linked to tumour progression. This supports the notion that the membranous evaluation of these proteins assessed by immunohistochemistry may be representative of their cell adhesion rather than their intra-cellular functions.     

Co-expression of stem cell genes CD133 and CD44 in colorectal cancers with early liver metastasis

PurposeTo investigate the expression status and clinical implications of stem cell genes CD133 and CD44 in the colorectal cancers with early liver metastasis.Materials and methodsThe differential genes of early liver metastases in colorectal cancer were detected by RT² Profiler? PCR Array. The expression and the relationship of stem cell gene CD133 and CD44 were analyzed by immunofluorescent tests.ResultsCD133 and CD44 were significantly higher co-expressed in colorectal cancer with early liver metastases compared to those without early liver metastases, and the content of CD133 and CD44 proteins decreased following growth of the transplanted tumors. Of the 80 cases without early liver metastases, 12 were observed CD133 and CD44 proteins co-expression, while 36 of the 40 cases with early liver metastases were found CD133 and CD44 proteins co-expression (15% vs. 90%, P < 0.05). Survival analysis revealed CD133 and CD44 proteins co-expression was associated with poorest prognosis (57.14% vs. 87.41%, X² = 48.49, P = 0.001). After Cox regression, age, Duck’s stage, lymph node metastasis, and CD133 and CD44 proteins co-expression were shown to be the independent prognostic factors of colorectal cancers.ConclusionsCD133 and CD44 proteins were highly co-expressed in colorectal cancer with early liver metastases, and may be a potential biomarker for the early liver metastases.     

肿瘤干细胞标志物CD133、CD44、OCT-4在小细胞肺癌中的表达及其临床意义

目的：探讨肿瘤干细胞标志物CD133、CD44、OCT-4与小细胞肺癌临床病理特征之间的相关性及临床意义。方法应用免疫荧光技术检测小细胞肺癌细胞株NCI-H82中肿瘤干细胞标志物CD133、CD44、OCT-4的表达;同时应用免疫组织化学法检测79例小细胞肺癌组织中CD133、CD44、OCT-4的表达。结果在小细胞肺癌细胞株NCI-H82中,CD133和CD 44的荧光信号为阳性表达,OCT-4的荧光信号为阴性表达。小细胞肺癌组织中CD133和CD44表达与肿瘤直径、淋巴结转移和临床分期相关,差异具有统计学意义（P＜0.05）。小细胞肺癌组织中OCT-4为阴性表达。结论 CD133和CD44可能是小细胞肺癌肿瘤干细胞的标志物,对小细胞肺癌的诊断和治疗有一定的临床意义。     

Clinicopathological significance of cancer stem cell markers CD44 and ALDH1 expression in breast cancer

Background

CD44 and aldehyde dehydrogenase 1 (ALDH1) has been reputed to be cancer stem cell (CSC) markers in breast cancer. Yet, the clinicopathologic and prognostic significance of these markers remain unclear. In this study, we have investigated the expression of these markers and their relation with conventional clinicopathologic tumor characteristic including molecular subtype.

Methods

CD44 and ALDH1 expression were investigated by immunohistochemistry in a series of 157 formalin-fixed paraffin-embedded breast cancer tissues.

Results

Overall, CD44 and ALDH1 are, respectively, detected in 33% (52 of 157) and 7% (10 of 157) of breast cancer cases. We also observed that CD44 expression was associated with histological grade (p?=?0.005). For ALDH1, we found that its expression is more frequent with elderly women (>?50 years, p?=?0.03). The investigation of relationship between the stem cell phenotype and breast cancer molecular subtype, revealed that CD44 and ALDH1 expression was more frequent in basal-like tumors (p?=?0.005). Among the two cancer stem cell markers tested, ALDH1 showed a strong association with the basal marker EGFR (p?=?0.05).

Conclusions

These findings suggest that CD44 and ALDH1 play a role in the clinical behavior in breast cancer and might be interesting biomarkers and therapeutic targets.

     

RNA aptamers targeting cancer stem cell marker CD133

The monoclonal antibody against the AC133 epitope of CD133 has been widely used as a cell surface marker of cancer stem cells in several different cancer types. Here, we describe the isolation and characterisation of two RNA aptamers, including the smallest described 15 nucleotide RNA aptamer, which specifically recognise the AC133 epitope and the CD133 protein with high sensitivity. As well, both these aptamers show superior tumour penetration and retention when compared to the AC133 antibody in a 3-D tumour sphere model. These novel CD133 aptamers will aid future development of cancer stem cell targeted therapeutics and molecular imaging.     

肿瘤干细胞标志物在结直肠癌中的研究进展

肿瘤干细胞(CSC)标志物是用来标记并鉴定CSC的一类分子.目前研究发现与结直肠癌有关的CSC标志物主要有CD133、CD29、CD166、CD44、Nanog等.这些干细胞标志物通过多种分子机制参与肿瘤的发生发展,可作为潜在的治疗靶点,还可被用作评估患者预后的有效指标.     

肿瘤干细胞及其与CD133的关系

越来越多的证据支持肿瘤干细胞理论,肿瘤可以认为是一个被少数肿瘤干细胞驱动的异常器官,它们具有自我更新和多向分化潜能.CD133在许多肿瘤中相继被报道,尽管其生物学功能还不清楚,但目前已经是一个非常有价值的分选肿瘤干细胞的标志,研究发现CD133+肿瘤细胞与肿瘤发生、侵袭、转移、耐药及复发有着密切的关系,因此深入了解CD133+肿瘤细胞的分子生物学特性对寻求有效的抗癌治疗,特别是靶向肿瘤干细胞的治疗是相当必要的.     

Brca1 breast tumors contain distinct CD44+/CD24- and CD133+ cells with cancer stem cell characteristics

Introduction

Whether cancer stem cells occur in BRCA1-associated breast cancer and contribute to therapeutic response is not known.

Methods

We generated and characterized 16 cell lines from five distinct Brca1deficient mouse mammary tumors with respect to their cancer stem cell characteristics.

Results

All cell lines derived from one tumor included increased numbers of CD44⁺/CD24^- cells, which were previously identified as human breast cancer stem cells. All cell lines derived from another mammary tumor exhibited low levels of CD44⁺/CD24^- cells, but they harbored 2% to 5.9% CD133⁺ cells, which were previously associated with cancer stem cells in other human and murine tumors. When plated in the absence of attachment without presorting, only those cell lines that were enriched in either stem cell marker formed spheroids, which were further enriched in cells expressing the respective cancer stem cell marker. In contrast, cells sorted for CD44⁺/CD24^- or CD133⁺ markers lost their stem cell phenotype when cultured in monolayers. As few as 50 to 100 CD44⁺/CD24^- or CD133⁺ sorted cells rapidly formed tumors in nonobese diabetic/severe combined immunodeficient mice, whereas 50-fold to 100-fold higher numbers of parental or stem cell depleted cells were required to form few, slow-growing tumors. Expression of stem cell associated genes, including Oct4, Notch1, Aldh1, Fgfr1, and Sox1, was increased in CD44⁺/CD24^- and CD133⁺ cells. In addition, cells sorted for cancer stem cell markers and spheroid-forming cells were significantly more resistant to DNA-damaging drugs than were parental or stem cell depleted populations, and they were sensitized to the drugs by the heat shock protein-90 inhibitor 17-DMAG (17-dimethylaminoethylamino-17-demethoxygeldanamycin hydrochloride).

Conclusion

Brca1-deficient mouse mammary tumors harbor heterogeneous cancer stem cell populations, and CD44⁺/CD24^- cells represent a population that correlates with human breast cancer stem cells.     

肿瘤干细胞标志物CD133-2、CD24和CD44S在头颈部鳞状细胞癌组织中的表达及其临床意义

 目的　探讨肿瘤干细胞标志物CD133-2、CD24、CD44S在头颈部鳞状细胞癌（HNSCC）组织中的表达情况及其临床意义。方法　采用免疫组织化学SP法,分别检测83例HNSCC患者癌组织、46例癌旁正常鳞状上皮组织中CD133-2、CD24、CD44S的表达,分析表达情况及与临床病理特征的关系。结果　CD133-2在癌组织、癌旁正常鳞状上皮组织中表达率分别为9.64 %（8／83）、21.74 %（10／46）,CD24分别为90.36 %（75／83）、45.65 %（21／46）,差异均有统计学意义（χ2值分别为15.040、5.818,均P＜0.05）;CD44S在癌组织、癌旁正常组织中均表达,但其染色评分差异有统计学意义（Z＝-4.262,P＜0.05）。在癌组织中,CD133-2的表达与组织的分化程度呈负相关（χ2＝7.246,P＜0.05）,CD24和CD44S的表达均与组织的分化程度呈正相关（χ2值分别为9.005、44.765,均P＜0.05）,CD44S的表达与T分期呈负相关（χ2＝4.650,P＜0.05）。结论　CD24、CD44S在HNSCC中的表达随着肿瘤细胞的分化程度增高而增高,能否作为HNSCC的肿瘤干细胞标志物尚需进一步研究;CD133-2表达随肿瘤细胞的分化程度增高而呈下调趋势,可以作为肿瘤干细胞的标志物,并可能与肿瘤的进展、临床预后相关。     

结直肠癌肿瘤干细胞标志物的研究进展

肿瘤干细胞与恶性肿瘤复发、耐药等生物学过程密切相关,是治疗恶性肿瘤的新靶点.寻找稳定性高、特异性强、实用性好的肿瘤干细胞标志物,清除恶性肿瘤中的肿瘤干细胞有望彻底根治恶性肿瘤.本文综述了结直肠癌中比较公认的肿瘤干细胞标志物CD44和潜在的肿瘤干细胞标志物LGR5、Bmi1、SOX2、Nanog的研究进展,为靶向治疗结直肠癌提供了参考依据.     

结直肠癌肿瘤干细胞标志物的研究进展

近年来,肿瘤干细胞（CSC）学说得到越来越多的支持,识别CSC的关键就是寻找干细胞的表面特异性标志物.结直肠癌CSC的相关研究不断取得进展,文章综述了近几年发现的比较公认的干细胞标志物CD44、SOX9、ALDH1和OCT-4等的研究进展及其与结直肠癌的关系.     

肿瘤干细胞表面标志物CD133单链抗体基因的分离与鉴定

 【摘要】　目的　分离、构建和表达抗人CD133单链抗体（scFv CD133）,测定其生物学活性。方法　用抗体工程技术从抗人CD133单克隆抗体（mAb）杂交瘤细胞中分离抗体可变区基因（VL和VH）,测定DNA序列并确定抗体互补决定区（CDR）;将scFv CD133基因克隆至pET32a中,转化Origami菌株,IPTG诱导,Ni2+-NTA His树脂纯化单链抗体,梯度硫氰酸盐洗脱法和酶联免疫吸附（ELISA）法检测其亲和性和特异性。结果　scFv CD133 基因VL和VH长度分别为339 bp和342 bp,各编码113和114个氨基酸,其中VL隶属于小鼠Igκ轻链,VH隶属于小鼠Ig重链I亚类。scFv CD133经SDS-PAGE和Western blot分析证明相对分子质量为27×103,体外实验具有一定的亲和性和特异性。结论　获得scFv CD133基因,为CD+133肿瘤干细胞的靶向治疗奠定了基础。     

肿瘤干细胞表面标志物CD133单链抗体基因的分离与鉴定

Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.     

肿瘤干细胞表面标志物CD133单链抗体基因的分离与鉴定

Objective To identify,construct and express scFv CD133,verify its biological function.Methods VL and VH were isolated from hybridoma of mAb CD133 by using antibody engineering technology.Its DNA sequencing and CDR were determined.scFv CD133 was then cloned into pET32a,transformed into Origami,induced by IPTG,purified by Ni2+-NTA His resin.Its affinity and specificity were tested by NH4SCN elution and ELISA.Results The size of VL and VH of scFv CD133 was 339 bp and 342 bp,which coded 113 and 114 amino acid separately.Its VL belonged to mouse Igκ chain and VH belonged to mouse IgG heavy chain subtype I.The molecular weight of scFv CD133 was about 27 × 103 which was testified by SDSPAGE and Western blot.Its affinity and specificity were also verified.Conclusion scFv CD133 has been successfully constructed and expressed in Origami,which could supply basis for target therapy of CD+133 cancer stem cell.