Journal: Experimental and Therapeutic Medicine

Article Title: Dual-targeting nanomicelles with CD133 and CD44 aptamers for enhanced delivery of gefitinib to two populations of lung cancer-initiating cells

doi: 10.3892/etm.2019.8220

Figure Lengend Snippet: Analysis of CD44 and CD133 expression in lung cancer cells and in vitro targeting of fluorescent nanomicelles. (A) Analysis of CD44 and CD133 expression in H446 cells. (B) Analysis of CD44 and CD133 expression in A549 cells. (C) In vitro targeting of fluorescent nanomicelles in (C) H446 CD133 + and CD133 − cells, (D) H446 CD44 + and CD44 − cells, (E) A549 CD133 + and CD133 − cells and (F) A549 CD44 + and CD44 − cells. Values are expressed as the mean ± standard deviation (n=3). *P<0.05. CD133-NM-Gef, CD133 aptamer-conjugated gefitinib 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-polyethylene glycol-2000 nanomicelles; PECF, 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-carboxyfluorescein.

Article Snippet: The cells were then incubated with fluorescent antibody (phycoerythrin-labeled CD133 antibodies; cat. no. FAB11331P-025; and Alexa Fluor ® 488-labeled CD44 antibodies; cat. no. FAB6127G; R&D Systems, Inc.) at a final concentration of 1 μg/ml on ice in a refrigerator.

Techniques: Expressing, In Vitro, Standard Deviation

Journal: Therapeutic Advances in Medical Oncology

Article Title: Association of early changes of circulating cancer stem-like cells with survival among patients with metastatic breast cancer

doi: 10.1177/17588359221110182

Figure Lengend Snippet: Demonstration of CD133 expression in circulating tumor cells (CTCs). The experimental control using HCT-116 [a human colon cancer cell line, CD133 + EpCAM + Hoechst + , (a)] and OECM-1 [a head and neck cancer cell line, CD133 − EpCAM + Hoechst + , (b)] by immunofluorescence staining. We further demonstrate a real breast cancer patient in the study cohort with CD133 + CTCs (c) and CD133 − CTC (d).

Article Snippet: Thereafter, we spiked the immunomagnetically enriched samples with OECM1/HCT116 cells labeled with an Alexa Fluor 488-conjugated monoclonal antibody to EpCAM (1:400; Cell Signaling Technology, Danvers, MA, USA) and an Alexa Fluor 647-conjugated monoclonal antibody to CD133 (1:200; Novus Biologicals, Littleton, CO, USA), and we stained the samples with Hoechst 33342 (20 mM; Thermo Fisher Scientific, Waltham, MA, USA), a blue fluorescent stain specific to DNA.

Techniques: Expressing, Control, Immunofluorescence, Staining

Journal: Journal of Experimental & Clinical Cancer Research : CR

Article Title: A tumor suppressor protein encoded by circKEAP1 inhibits osteosarcoma cell stemness and metastasis by promoting vimentin proteasome degradation and activating anti-tumor immunity

doi: 10.1186/s13046-024-02971-7

Figure Lengend Snippet: KEAP1-259aa exerts the biological function in OS cells. ( A ). OS cells were transfected with empty vector, circKEAP1 vector, ATG-mutated circKEAP1 vector and IRES-mutated circKEAP1 vector. Colony formation was performed in the indicated transfection groups. ( B ). Cell proliferation of the indicated cells was characterized by CCK-8 assay. ( C ). Apoptosis of the indicated cells was detected by flow cytometry. ( D ). Cell apoptosis of the indicated cells was characterized by western blotting assay. ( E ). Cell migration of the indicated cells was characterized by wound healing assay. ( F ). Cell migration of the indicated cells was characterized by transwell assay. ( G ). Tumor sphere assay was performed to detect stemness of the indicated cells (scale bar, 50 μm). ( H ). Flow cytometry assay was used to examine percentage of CD133 + cells in the indicated cells. ( I ). SOX2, ABCG2, Oct4 and Nanog expression in cells was characterized by western blotting. ( J ). The indicated cells (1 = NC, 2 = KEAP1-259aa, 3 = IRSE mut, 4 = ATG mut) were injected subcutaneously into nude mice ( n = 4 per group) and tumor volume was measured every 7 days. After 28 days, tumors were collected and weighed. ( K ). The indicated cells were injected into nude mice ( n = 4 per group) via tail vein and the lungs were harvested after 35 days. Representative image and H&E staining of mouse lung metastases is shown (scale bars, 200 μm). Error bars represent three independent experiments. *, **, *** indicates significant differences compared with the control group at a p value < 0.05, < 0.01, < 0.001, respectively

Article Snippet: Human CD133 Alexa Fluor 700-conjugated antibody was obtained from RD Systems (MN, USA).

Techniques: Transfection, Plasmid Preparation, CCK-8 Assay, Flow Cytometry, Western Blot, Migration, Wound Healing Assay, Transwell Assay, Expressing, Injection, Staining, Control

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: Flow cytometry graphs of cluster of differentiation 133 expression in Tca-8113 cells. Results from four separate experiments are presented. As shown, 0.95% of the cells were highly expressed on the cell membrane. PE, phycoerythrin.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: Flow Cytometry, Expressing, Membrane

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: CD133 + cells stained with a green fluorescent protein observed with a fluorescence microscope (fluorescein isothiocyanate; ×200 magnification). Green fluorescence exhibited a scattered distribution, which indicated that CD133 was distributed on the cell membranes. Only a low percentage of whole-tongue squamous Tca8113 carcinoma cells expressed CD133. CD133, cluster of differentiation 133.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: Staining, Fluorescence, Microscopy

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: CD133 + cells purified by MACS. Prior to MACS, 1.24% of the Tca-8113 cells expressed CD133. After cell sorting, 92.45% of the Tca-8113 cells expressed CD133. Red indicates criteria curves; green and blue indicate the ratios of the cells prior to and after sorting, respectively. CD133, cluster of differentiation 133; MACS, magnetic-activated cell sorting.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: Purification, FACS

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: Proliferation states for CD133 + , CD133 – and primary tumour cells. CD133 + cells grew quickly between days 3 and 5; CD133 – cells grew much more slowly, indicating less proliferative activity. CD133, cluster of differentiation 133; OD, optical density.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: Activity Assay

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: Differentiation curve of CD133 + cells in vitro . During 16 days of culture, the percentage of CD133 + cells decreased from 92.45 to 1.62%. CD133, cluster of differentiation 133.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: In Vitro

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: Comparison of the tumourigenic capacity of CD133 + , CD133 – and primary cells in NOD/SCID mice. The subcutaneous tumours (A) prior to and (B) after the NOD/SCID mice were sacrificed. (C) The tumours generated from the CD133 + cells were significantly larger compared with the tumours generated from the CD133 – cells. In addition, in terms of the tumourigenic time, CD133 + cells could generate tumours in ~1 week, whereas CD133 – cells required 10 to 14 days or longer.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: Comparison, Generated

Journal: Oncology Letters

Article Title: Role of CD133 + cells in tongue squamous carcinomas: Characteristics of ‘stemness’ in vivo and in vitro

doi: 10.3892/ol.2016.4719

Figure Lengend Snippet: Comparison of the number of cancers generated from different cell types.

Article Snippet: Next, 30 μl fluorescein isothiocyanate (FITC)-labelled anti-CD133 antibody (catalog no. 46133182; R&D Systems Inc., Minneapolis, MN, USA) was added to cover the cells, which were incubated for 1 h at room temperature in the dark.

Techniques: Comparison, Generated

Journal: Stem Cell Reviews

Article Title: Endothelial Progenitor Cells Modulate Inflammation-Associated Stroke Vasculome

doi: 10.1007/s12015-019-9873-x

Figure Lengend Snippet: EPC treatment downregulates the OGD-induced upregulation of inflammation-associated stroke vasculome in vitro in HEN6 cells and increases cell viability. a Representative photomicrographs of human bone marrow mesenchymal stem cells (BM-MSCs) seeded with EGM-2MV medium supplemented with growth factors on collagen-I coated 48 well plates and incubated for 7 days. After 7 days, immunocytochemistry analysis revealed specific expression of EPC markers including vWF, CD133 and VEGFR-2. Next, human endothelial brain cells (HEN6) were subjected to oxygen and glucose degradation (OGD) and co-cultured with EPCs and levels of RNA vasculome were analyzed with qRT-PCR. b Quantitative analyses of total fold change of RNA vasculome revealed that BRM, IκB, Foxf1, and ITIH-5 levels increased after OGD up to 10 folds relative to ambient conditions (Two way ANOVA, F 3,21 = 19.86, p < 0.001, Bonferonni’s test, p < 0.05). After EPC treatment, there was a significant 4, 6, and 2 fold decrease expression of BRM, IkB, and Foxf1 respectively in OGD-EPC treated HEN6 cells relative to OGD-vehicle treated HEN6 cells (p’s < 0.05), except ITI-H5 ( p > 0.05). c Cell viability analysis showed there was a significant increase of HEN6 cell survival in OGD-EPC and OGD-EPC scramble treatment compared to OGD alone or OGD-EPC IkB knockdown (One way ANOVA, F 1,22 = 44.58, p < 0.001, Bonferonni’s test, p < 0.05)

Article Snippet: Three different endothelial progenitor cell markers were used including CD133 (1:100, Catalog# NBP2–37741, Novus Biologicals), anti-VEGF (1:100, Catalog# ab2349, Abcam) and anti-von Willebrand factor antibody (1:100, Catalog# MAB3442, Merck Millipore).

Techniques: In Vitro, Incubation, Immunocytochemistry, Expressing, Cell Culture, Quantitative RT-PCR, Knockdown