Expression data from HCT116 cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
SUBMITTER: Alexandra Graf
PROVIDER: E-GEOD-38049 | biostudies-arrayexpress |
REPOSITORIES: biostudies-arrayexpress
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Project description:Advanced colon cancer is characterized by drug resistance and a poor prognosis. In these patients tumor-propagating cells appear to be largely resistant against various targeted drugs including ErbB-inhibitors. The cell surface antigen prominin-1 (CD133) has recently been identified as a potential marker of colon cancer stem cells. The purpose of this study was to define mRNA expression patterns in CD133+ and CD133- HCT116 cells.
2012-05-22 | GSE38049 | GEO
Project description:BACKGROUND: Several in vitro assays have been used to identify “cancer stem cells” (CSC), including expression of cell surface markers and Hoechst dye efflux properties. However, each of these methods has potential pitfalls that complicate interpretation of the results. Focusing on colon cancers (CC), the CD133 antigen has been proposed as a marker of colon CSC. However, conflicting results have been reported in the literature indicating the need of a systematic analysis of CSC within CC and a complete validation of markers for the isolation of these cells. AIMS: Aim of this study was to confirm that CD133 expression is a valid method for isolating CSC in CC and verify if other antigens can increase the specificity of this marker for isolating CSC in CC. METHODS: CD133+ and CD133- cells were isolated from different human CC lines (CaCo-2, HT29, LOVO, HCT-116) by FACS sorter and the tumor-initiating potential of CD133+ cells was assessed in vitro, by soft-agar colony formation assay, and in vivo, upon transplantation into nude mice. Furthermore, the gene expression profile of CD133+ versus CD133- CaCo-2 cells was compared by the means of microarray analysis. Then, in the effort to identify a common “tumor stem cell” signature for CC, the most relevant transcripts resulting from gene expression profiling on CD133+ cells was assessed by real-time PCR on SP-fraction isolated by FACS sorter from the same CC cell lines. Finally, we deplete CD133 expression in the CaCo-2 cell line by the means of siRNA and verified by Western Blot analysis whether there was a functional correlation between CD133 and the target genes. Moreover, CaCo-2 and HCT116 cells were exposed to sodium butyrate (NaBu) for 72h. Colon cells differentiation was assessed by Alkaline phosphatase activity and expression of CD133 and target genes was tested by western blot. RESULTS: We confirmed that only CD133+ cells have a tumor-initiating potential in vitro and in vivo. Furthermore, microarray analysis of CD133+ versus CD133- CaCo-2 cells revealed a significant overexpression of various transcripts involved in cell proliferation, invasion and stemness in CD133+ cell fraction. Comparison of the transcripts by real-time PCR revealed that the genes of Endothelin-1 (END-1) and NR4A2 are highly expressed in both CD133 + cells and in SP fractions. Finally, when we deplete CD133 expression in Caco-2cells by siRNA, we observed a significant attenuation of END-1 and NR4A2 expression, thus demonstrating that CD133 is involved in the transcriptional regulation of these genes. Interestingly, we also showed that the expression of all three genes was inversely correlated with cell differentiation status as demonstrated by the fact that their expression decreases in a time- and dose-dependent manner after differentiation induced by NaBu. CONCLUSION: Overall, this study confirms the role of CD133antigen as CSC marker and showed for the first time the existence of a functional relationship between CD133, END-1 and NR4A2 expression, hypothesizing that CD133 is involved in the transcriptional regulation of these gene. Microarray analysis was performed on CD133+ and CD133- sorted CACO-2 cells. For both fractions, cells were sorted three independent times. Sample preparation was performed according to Affymetrix recommendations. A total of 6 arrays were hybridized, including 3 CD133+ replicates and 3 CD133- replicates.
2011-01-04 | E-GEOD-24747 | biostudies-arrayexpress
Project description:CD133 has been widely used for identification and isolation of cancer stem cells in tumors although its role as a marker for cancer stem cell is still controversial . We isolated the CD133+ and CD133- cells from SW620 human colon cancer cell line and compared their biological characteristics, such as tumorigenicity,drug sensitivity, etc. Our study revealed that CD133+ SW620 cells were more tumorigenic and resistant to anti-cancer drugs. Correspondingly, they displayed different gene expression profile. However, it was observed that CD133- cells and CD133+ cells could mutually convert, indicating that CD133 expression was under dynamic and reversible regulations which might impose significant infulence on cells behaviors. Thus, our data challenge the role of CD133 as a marker for cancer stem cell. There are two populations with distinct expression of CD133 in SW620 human colon cancer cell line. Microarray assays were employed to investigate the differentially expressed genes between the two populations, which may possess different tumorigenetic potential and sensitivity to anti-cancer drugs. CD133+ and CD133- cells were isolated from human colon cancer SW620 cell line by magnetic cell sorting system. The clones from sorted CD133+ or CD133- populations were established. Clone cells were expanded and were further purified by using CD133 cell isolation kit before microarray assays.
2010-07-29 | E-GEOD-23295 | biostudies-arrayexpress
Project description:Expression from CD133+ cells isolated from adult human exocrine tissue was compared to a CD133-depleted cell population Islet-depleted exocrine tissue from three independent adult human cadaveric pancreata were cultured for four days in Miami media 1A. Following trypsinization, cells were isolated using anti-CD133 immunomagnetic beads to >95% CD133+. CD133-negative cells were further depleted of CD133+ cells to <1% CD133+.
2016-01-15 | E-GEOD-64854 | biostudies-arrayexpress
Project description:Transcriptional profiling of GIF-5 mouse gastric epithelial cells comparing CD133-positive and CD133-negative cells. The former formed CD133-positive and CD133-negative cells while the latter only CD133-negative cells, suggesting that CD133-positive cells are mother cells. The former produced differentiated type tumors while the latter undifferentiated types in vivo, indicating a relationship between CD133-expression and glandular structure formation. One-condition experiment, CD133-positive vs. CD133-negative cells. 2 replicates.
2011-08-04 | E-GEOD-23456 | biostudies-arrayexpress
Project description:Cancer stem cells (CSCs) that display tumor-initiating properties have recently been identified. We herein identify and characterize CSCs in human uterine carcinosarcoma, a highly aggressive and therapy-resistant gynecologic malignancy, which is considered to be of mesodermal origin. FU-MMT-1, a cell-line, which was established by us (Emoto M, Cancer 1992) from a patient with uterine carcinosarcoma, was evaluated. FU-MMT-1 contained a high population of CD133, CD44, CD90, and CD29 positive cells. Using the magnetic bead cell separation method, we isolated CD133+ cells, which predominantly form spheres in culture. These CD133+ cells form transplantable tumors in vivo. A qRT-PCR analysis of the genes implicated in stem cell maintenance revealed that CD133+ cells express significantly higher levels of OCT4, NANOG, and BMI-1 than CD133M-oM-<M- cells. Moreover, CD133+ cells showed a high expression of PAX2 and WNT4, which are the essential genes in Mullerian duct formation. The tumor derived from CD133+ cells replicated vimentin, ERM-NM-1, ERM-NM-2, and PR expressionsM-cM-^@M-^@of the parent tumor. These findings suggest that CD133+ FU-MMT-1 cells have the characteristics of CSCs and Mullerian mesenchymal progenitors. CD133+ and CD133- population of FU-MMT-1 cells were analyzed by microarray.
2013-02-10 | E-GEOD-27172 | biostudies-arrayexpress
Project description:To identify regulators of human HSC fate, we transcriptionally profiled quiescent primitive cord blood (CB) CD133+ G0 cells enriched for long term culture initiating cell (LTC-IC) activity. CD133+ G0 cells were sorted by cell cycle status using combined DNA and RNA staining; less immature CD133+G1 cells served as a comparison.
2008-03-06 | E-MEXP-1412 | biostudies-arrayexpress
Project description:Overarching aim is to achieve a greater understanding of the control of progenitor cells within the adult human retina within the normal and diseased retinal microenvironment. Specifically we will assess via our experimental designs: (i) the control of CD133+ retinal cell populations that display mitotic potential and differentiation and Keywords: comparative genomic hybridization using adult human CD133+ & CD133- retinal cells Adult human retina was processed to produce a cell suspension, CD133+ cells were magnetically isolated to produce cell populations of CD133+ and CD133- cells. 3 retinal samples were analysed for each cell population.
2009-02-06 | E-GEOD-14733 | biostudies-arrayexpress
Project description:Nine heterogeneous melanoma cell lines including D10 and WM115 were studied for cancer stem cell characteristics in vitro. D10 cell line was the only cell line expressing the cancer stem cell marker CD133. Thus, gene expression profiling on CD133+ and CD133- D10 cells was carried out using Affymetrix GeneChips Human Genome U133A 2.0.
2013-01-17 | E-MEXP-3542 | biostudies-arrayexpress
Project description:As many other tumors, a subset of gliobastoma is thought to be maintained by a restricted population of cancer cells, stem-like cells that express CD133 transmembrane protein. Expression levels of CD133 gene has been linked to a poor prognostic molecular subgroup and is not overexpressed by the PDGF-driven proneural group. Thus, the significance of CD133+ cells for gliomagenesis of the proneural group is undetermined. In addition, the role of the CD133 protein remains elusive and controversial, which results from the difficult isolation of CD133+ cells that has largely relied on the use of antibodies to ill-defined glycosylated epitopes of CD133. Here, we used a knockin lacZ reporter mouse, Prom1lacZ/+, to track Prom1+ cells in the brain and found that Prom1 (prominin1, murine CD133 homologue) is expressed by cells that co-express markers characteristic of neuronal, glial and vascular lineage phenotype. In proneural tumors derived from injection of RCAS-PDGF into the brain of tv-a;Ink4a-Arf-/- Prom1lacZ/+ mice, Prom1+ cells co-express markers for astrocytes and endothelial cells. Therefore, we characterize the tumor propagation in a murine model and found that the mice co-transplanted with Prom1 endothelium and proneural tumor spheres cells had significant tumor burden and vascular proliferation (angiogenesis). Specific genes in Prom1 endothelium are identified that code for endothelial signaling modulators that most likely support proneural tumor progression and can be potential targets for anti-angiogenic therapy. Cells were sorted via FACS to obtain a population of CD31+CD133- cells and a population of CD31+CD133+ cells. Total RNA was extracted from each population and gene expression was assayed on Affymetrix Mouse 430 2.0 arrays with one array per cell population.
2013-12-31 | E-GEOD-43931 | biostudies-arrayexpress