Journal: Oncology Reports

Article Title: Hypoxia induces radioresistance, epithelial-mesenchymal transition, cancer stem cell-like phenotype and changes in genes possessing multiple biological functions in head and neck squamous cell carcinoma

doi: 10.3892/or.2022.8269

Figure Lengend Snippet: Relative mRNA expression of epithelial-mesenchymal transition and cancer stem cell markers in hypoxic head and neck squamous cell carcinoma cells.

Article Snippet: The expression of hypoxia-responsive genes was analyzed with a panel of TaqMan ® Gene Expression assays [cadherin 1 (CDH1; Hs01023895_m1); cadherin 2 ( CDH2 ; Hs00983056_m1); vimentin (VIM; Hs00958111_m1); fibronectin 1 (FN1; Hs01549976_m1); FOXC2 (Hs00270951_s1); TWIST1 (Hs04989912_s1); CD44 (Hs01075864_m1); SOX2 (Hs04234836_s1); NANOG (Hs02387400_g1); GLUT3 (Hs00359840_m1); CA9 (Hs00154208_m1); caspase 14 ( CASP14 ; Hs00201637_m1); serpin family E 1 ( SERPINE1 ; Hs00167155_m1); lysyl oxidase ( LOX ; Hs00942480_m1); amphiregulin ( AREG ; Hs00950669_m1); epiregulin ( EREG ; Hs00914313_m1); all labelled with FAM and purchased from Thermo Fisher Scientific, Inc.] and amplified using a TaqMan real-time PCR protocol according to manufacturer's instructions (Thermo Fisher Scientific, Inc.).

Techniques: Expressing

Journal: Stem Cells International

Article Title: BMI1 , ALDH1A1 , and CD133 Transcripts Connect Epithelial-Mesenchymal Transition to Cancer Stem Cells in Lung Carcinoma

doi: 10.1155/2016/9714315

Figure Lengend Snippet: mRNA expression levels of EMT ( BMI1, TWIST1 ), CSC ( CD133, ALDH1A1 ), and epithelial ( EpCAM ) markers in primary tumors and whole blood samples of 57 NSCLC patients. Data are shown as scattered plot for all measured values with line in the middle representing the median.

Article Snippet: The TaqMan assays EpCAM (CD326) (Hs00901885_m1), BMI1 (Hs00180411_m1), TWIST1 (Hs00361186_m1), PROM1 (CD133) (Hs1009245_m1), and ALDH1A1 (Hs00167445_m1) were used.

Techniques: Expressing

Journal: Stem Cells International

Article Title: BMI1 , ALDH1A1 , and CD133 Transcripts Connect Epithelial-Mesenchymal Transition to Cancer Stem Cells in Lung Carcinoma

doi: 10.1155/2016/9714315

Figure Lengend Snippet: Spearman's rank correlation coefficient analysis of associations between mRNA expression of EMT ( BMI1 , TWIST1 ), CSC ( CD133 , ALDH1A1 ), and epithelial ( EpCAM ) markers in primary tumors and matched whole blood samples of 57 operable NSCLC patients. Statistically significant associations ( P < 0.01) between EMT and CSC marker expression are presented in bold.

Article Snippet: The TaqMan assays EpCAM (CD326) (Hs00901885_m1), BMI1 (Hs00180411_m1), TWIST1 (Hs00361186_m1), PROM1 (CD133) (Hs1009245_m1), and ALDH1A1 (Hs00167445_m1) were used.

Techniques: Expressing, Marker

Journal: Stem Cells International

Article Title: BMI1 , ALDH1A1 , and CD133 Transcripts Connect Epithelial-Mesenchymal Transition to Cancer Stem Cells in Lung Carcinoma

doi: 10.1155/2016/9714315

Figure Lengend Snippet: Expression levels of CSC ( CD133, ALDH1A1 ) and EMT ( BMI1, TWIST1 ) in EpCAM-positive CTCs of 10 patients with metastatic NSCLC. Dashed line represents relative mRNA level in PBMNCs. ALDH1A1 and BMI1 transcripts were detected in 10/10 patients whereas CD133 and TWIST1 transcripts were detected in 5/10 and 4/10 patients, respectively. Data are shown as scattered plot for all measured values with line in the middle representing the median mRNA expression level of marker in CTCs.

Article Snippet: The TaqMan assays EpCAM (CD326) (Hs00901885_m1), BMI1 (Hs00180411_m1), TWIST1 (Hs00361186_m1), PROM1 (CD133) (Hs1009245_m1), and ALDH1A1 (Hs00167445_m1) were used.

Techniques: Expressing, Marker

Journal: Blood

Article Title: The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

doi: 10.1182/blood-2009-09-242313

Figure Lengend Snippet: Patient and disease characteristics and TWIST levels in CD34 + cells

Article Snippet: The membranes were blocked in 5% nonfat dry milk diluted in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hour at room temperature and then incubated overnight at 4°C in 5% nonfat dry milk /TBS-T containing either rabbit anti-P53 antibody (1:1000; Cell Signaling Technology Inc), rabbit anti-TWIST antibody (1:1000; Cell Signaling Inc), rabbit anti– intercellular adhesion molecule-1 (ICAM1) antibody (1:1000; Cell Signaling Inc) or rabbit anti-PYCARD antibody (1:1000; Sigma).

Techniques: Expressing

Journal: Blood

Article Title: The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

doi: 10.1182/blood-2009-09-242313

Figure Lengend Snippet: Expression of TWIST in patients with MDS and in healthy donors. (A) Stroma cells. mRNA levels were determined by quantitative real-time PCR. TWIST levels were lower in MDS than in healthy controls (see text). (B) CD34+ cells. Samples from 56 patients with MDS and from 17 healthy donors were analyzed. TWIST mRNA levels were significantly higher in patients with advanced MDS (P < .001). Shown are the means and SEM. The gel shows actual mRNA levels in 14 patients with MDS and 5 healthy controls. (C) TWIST levels in patients with MDS and healthy donors. Relative mRNA levels of TWIST were significantly higher in patients with low-grade (P < .001) and advanced MDS (P < .001) than in healthy donors. (D) Ratio of relative TWIST and p53 levels in patients with MDS and healthy donors. The ratio of TWIST to p53 mRNA differed significantly between cells from healthy donors and cells from patients with low-grade (P < .001) and advanced MDS (P < .001), respectively. Horizontal lines indicate median values.

Article Snippet: The membranes were blocked in 5% nonfat dry milk diluted in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hour at room temperature and then incubated overnight at 4°C in 5% nonfat dry milk /TBS-T containing either rabbit anti-P53 antibody (1:1000; Cell Signaling Technology Inc), rabbit anti-TWIST antibody (1:1000; Cell Signaling Inc), rabbit anti– intercellular adhesion molecule-1 (ICAM1) antibody (1:1000; Cell Signaling Inc) or rabbit anti-PYCARD antibody (1:1000; Sigma).

Techniques: Expressing, Real-time Polymerase Chain Reaction

Journal: Blood

Article Title: The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

doi: 10.1182/blood-2009-09-242313

Figure Lengend Snippet: TWIST expression and interactions with p53 in primary CD34+ marrow cells and myeloid cell lines. (A) Primary CD34+ marrow cells from healthy donors and patients with MDS. Healthy CD34+cells (control) cultured without stroma (Not-CC) or with stroma contact (CC) showed no change in the expression of p53 or TWIST. In cells from patients with MDS, p53 showed prominent up-regulation on stroma contact (CC), whereas levels of TWIST declined. (B) Immunoprecipitation (IP) of TWIST from KG1a, MDS-L, HL-60, and PL-21 cell lysates. p53 was contained in the complex precipitated from all myeloid cell lines by anti-TWIST antibody; DJ-1 served as positive (+) and Stro-1 as negative (−) control. (C) IP as in panel B, but using KG1a cells without and with coculture, normalizing for TWIST content in the cell lysate. IgG indicates immunoglobulin G.

Article Snippet: The membranes were blocked in 5% nonfat dry milk diluted in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hour at room temperature and then incubated overnight at 4°C in 5% nonfat dry milk /TBS-T containing either rabbit anti-P53 antibody (1:1000; Cell Signaling Technology Inc), rabbit anti-TWIST antibody (1:1000; Cell Signaling Inc), rabbit anti– intercellular adhesion molecule-1 (ICAM1) antibody (1:1000; Cell Signaling Inc) or rabbit anti-PYCARD antibody (1:1000; Sigma).

Techniques: Expressing, Control, Cell Culture, Immunoprecipitation, Negative Control

Journal: Blood

Article Title: The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

doi: 10.1182/blood-2009-09-242313

Figure Lengend Snippet: Inhibition of TWIST in KG1a cells and its effect on apoptosis. Early stage apoptosis (annexin V+, propidium iodide−) in KG1a and PL-21 cell lines transfected with either a scrambled siRNA sequence (SCR) or siRNA specific for TWIST (siTWIST), and exposed overnight to normal medium (veh) versus exogenous TNFα (25 ng/mL, A) or TRAIL (100 ng/mL, B), respectively. Apoptosis was determined by flow cytometry. (C) Cell preparation as in panels A and B, but cultured in normal medium (veh) or in the presence of the soluble TNF receptor etanercept (10 μg/mL for 24 hours [concentration based on ancillary studies]). Error bars indicate SEM. (D) Protein lysates from the same cell preparations were separated on 4% to 12% Bis-Tris gels and immunoblotted with antibodies against TWIST, p53-ser46, Bax, and caspase 9, respectively. β-Actin served as loading control. (E) TWIST and p53 levels in primary CD34+ MDS cells in which TWIST was silenced by specific siRNA (KD TWIST) compared with levels in unmodified cells from the same primary CD34+ MDS sample.

Article Snippet: The membranes were blocked in 5% nonfat dry milk diluted in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hour at room temperature and then incubated overnight at 4°C in 5% nonfat dry milk /TBS-T containing either rabbit anti-P53 antibody (1:1000; Cell Signaling Technology Inc), rabbit anti-TWIST antibody (1:1000; Cell Signaling Inc), rabbit anti– intercellular adhesion molecule-1 (ICAM1) antibody (1:1000; Cell Signaling Inc) or rabbit anti-PYCARD antibody (1:1000; Sigma).

Techniques: Inhibition, Transfection, Sequencing, Flow Cytometry, Cell Culture, Concentration Assay, Control

Journal: Blood

Article Title: The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

doi: 10.1182/blood-2009-09-242313

Figure Lengend Snippet: TWIST and p53 expression and apoptosis in primary CD34+ cells. (A) Apoptosis in CD34+ marrow cells from healthy normal bone marrow donors (NBM) and patients with MDS cocultured with either unmodified HS5 stroma or HS5 with suppression of TWIST by siRNA (KD TWIST). Apoptosis was determined by flow cytometry. Error bars indicate SEM. (B) Levels of TWIST and p53 mRNA in CD34+ cells from healthy donors (NBM) and patients with MDS cocultured with either unmodified HS5 stroma (WT) or HS5 stroma with inhibition of TWIST (KD TWIST).

Article Snippet: The membranes were blocked in 5% nonfat dry milk diluted in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hour at room temperature and then incubated overnight at 4°C in 5% nonfat dry milk /TBS-T containing either rabbit anti-P53 antibody (1:1000; Cell Signaling Technology Inc), rabbit anti-TWIST antibody (1:1000; Cell Signaling Inc), rabbit anti– intercellular adhesion molecule-1 (ICAM1) antibody (1:1000; Cell Signaling Inc) or rabbit anti-PYCARD antibody (1:1000; Sigma).

Techniques: Expressing, Flow Cytometry, Inhibition

Journal: Blood

Article Title: The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

doi: 10.1182/blood-2009-09-242313

Figure Lengend Snippet: Inhibition of TWIST and ICAM1 (CD54) expression in HS5 cells. (A) FACS analysis of ICAM1 expression in HS5 cells transfected with scrambled siRNA (SCR) or TWIST-specific siRNA (KD TWIST). The cells were stained with FITC conjugated anti–human CD54 antibody (AB) for 30 minutes (a indicates isotype control; b, HS5 transfected with scrambled siRNA sequences; c, HS5 transfected with TWIST-specific siRNA.) (B) Western blot of TWIST and ICAM1 proteins in HS5 cells transfected with scrambled siRNA (SCR) or with 1 of 3 TWIST-specific siRNAs (KD TWIST in HS5). β-Actin served as loading control. (C) IACM1 expression in primary MDS stroma cells; a indicates normal bone marrow stroma labeled with isotype control antibody; b, normal bone marrow stroma labeled with CD54+-allophycocyanin flow antibody; c1, patients with RA MDS; c2, patients with RAEB-2 MDS. (D) Early stage apoptosis (annexin V+/propidium iodide−) in KG1a cells in control cultures containing unmodified HS5 cells (WT) or HS5 cells pretreated with either 5 or 10 μg of anti-ICAM1 antibody. Cells were cultured in the absence or presence of TNFα (25 ng/mL). Apoptosis was determined by flow cytometry. Only CD45+ (KG1a) cells were considered. (E) Early-stage apoptosis in PL-21 and KG1a cells cultured with either unmodified stroma or stroma pretreated with anti-IACM1 antibody, and apoptosis in PL-21 or KG1a cells treated with anti-CD11b antibody. Error bars indicate SEM.

Article Snippet: The membranes were blocked in 5% nonfat dry milk diluted in Tris-buffered saline containing 0.1% Tween-20 (TBS-T) for 1 hour at room temperature and then incubated overnight at 4°C in 5% nonfat dry milk /TBS-T containing either rabbit anti-P53 antibody (1:1000; Cell Signaling Technology Inc), rabbit anti-TWIST antibody (1:1000; Cell Signaling Inc), rabbit anti– intercellular adhesion molecule-1 (ICAM1) antibody (1:1000; Cell Signaling Inc) or rabbit anti-PYCARD antibody (1:1000; Sigma).

Techniques: Inhibition, Expressing, Transfection, Staining, Control, Western Blot, Labeling, Cell Culture, Flow Cytometry

Journal: Oncology Reports

Article Title: Unravelling heterogeneous effects of cancer‑associated fibroblasts on poor prognosis markers in breast cancer EM‑G3 cell line: In vitro ‑targeted treatment (anti‑IL-6, anti‑VEGF-A, anti‑MFGE8) based on transcriptomic profiling

doi: 10.3892/or.2023.8662

Figure Lengend Snippet: Reagents used for western blot staining of cells.

Article Snippet: TWIST1 , Rabbit , IgG , Monoclonal , 1:1,000 , 69366 , Cell Signaling Technology, Inc..

Techniques: Western Blot, Staining, Produced

Journal: Oncology Reports

Article Title: Unravelling heterogeneous effects of cancer‑associated fibroblasts on poor prognosis markers in breast cancer EM‑G3 cell line: In vitro ‑targeted treatment (anti‑IL-6, anti‑VEGF-A, anti‑MFGE8) based on transcriptomic profiling

doi: 10.3892/or.2023.8662

Figure Lengend Snippet: Western blot analysis of EM-G3 cells. Protein expression analysis in indirect (CM) co-culture of EM-G3 cells with HDFs, BCCFs, SCCFs and BCMFs. The studied conditions for each studied fibroblast type included: Control culture of EM-G3 cells, EM-G3 cells cultured in CM derived from respective fibroblasts, EM-G3 cells cultured in CM enriched with neutralizing antibodies against either IL-6 or VEGF-A or MFGE8 or combination of all three tested antibodies (anti-IL-6 + anti-VEGF-A + anti-MFGE8). The expression profile of the following markers related to cell differentiation and epithelial-to-mesenchymal transition was evaluated: keratin-8, keratin-14, keratin-18, keratin-19, vimentin, SLUG, SNAIL, TWIST1, ZEB1, E-cadherin, N-cadherin, VE-cadherin. β-Actin was used as sample loading control (representative beta-actin controls are shown). HDFs, human dermal fibroblasts; BCCFs, basal cell carcinoma fibroblasts; SCCFs, squamous cell carcinoma fibroblasts; BCMFs, breast cancer metastasis fibroblasts; CM, conditioned media.

Article Snippet: TWIST1 , Rabbit , IgG , Monoclonal , 1:1,000 , 69366 , Cell Signaling Technology, Inc..

Techniques: Western Blot, Expressing, Co-Culture Assay, Control, Cell Culture, Derivative Assay, Cell Differentiation