Journal: International Journal of Molecular Medicine

Article Title: Downregulation of chemokine (C-C motif) ligand 5 induced by a novel 8-hydroxyquinoline derivative (91b1) suppresses tumor invasiveness in esophageal carcinoma

doi: 10.3892/ijmm.2024.5435

Figure Lengend Snippet: Compound 91b1 downregulates the expression of CCL5 in ESCC cell lines. CCL5 mRNA expression levels in ESCC cell lines (A) KYSE150, (B) KYSE450, (C) KYSE510 and (D) HKESC-4 after 48-h treatment of 91b1 in different concentrations. Each test was performed in triplicate and relative CCL5 expression levels were determined by comparing with cells treated with vehicle DMSO (0.05%) following normalized with the expression of β-actin. CCL5 protein expression of (E) KYSE150, (F) KYSE450, (G) KYSE510 and (H) HKESC-4 after 48-h treatment of 91b1 in different concentrations vs. vehicle. Each assay was performed in triplicate. (I) Immunohistochemical staining of CCL5 for KYSE150 treated with vehicle control (DMSO), 6.5, 9.5 and 21 μ g/ml 91b1 for 48 h; original magnification, ×400; (J) The staining signals were quantitatively analyzed by ImageJ (National Institutes of Health). * P<0.05; ** P<0.01; ns, not significant. CCL5, chemokine (C-C motif) ligand 5; ESCC, esophageal squamous cell carcinoma.

Article Snippet: Esophageal squamous cell carcinoma (ESCC) cell lines of Japanese origin KYSE150, KYSE450, KYSE30 and KYSE510 ( ) were purchased from Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH.

Techniques: Expressing, Immunohistochemical staining, Staining, Control

Journal: International Journal of Molecular Medicine

Article Title: Downregulation of chemokine (C-C motif) ligand 5 induced by a novel 8-hydroxyquinoline derivative (91b1) suppresses tumor invasiveness in esophageal carcinoma

doi: 10.3892/ijmm.2024.5435

Figure Lengend Snippet: Compound 91b1 shows stronger cytotoxic effect in ESCC cell lines and less cytotoxic effect in non-neoplastic cells. Cytotoxic effects of 91b1 on ESCC cell lines (A) KYSE30, (B) KYSE150, (C) KYSE450, and (D) HKESC-4; and non-neoplastic cell lines (E) NE-3 and (F) 293 were examined by MTS cytotoxicity assay with CDDP as the positive control. Each test was performed in triplicate. ESCC, esophageal squamous cell carcinoma; MTS, [3-(4,5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium]; CDDP, cisplatin.

Article Snippet: Esophageal squamous cell carcinoma (ESCC) cell lines of Japanese origin KYSE150, KYSE450, KYSE30 and KYSE510 ( ) were purchased from Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH.

Techniques: Cytotoxicity Assay, Positive Control

Journal: International Journal of Molecular Medicine

Article Title: Downregulation of chemokine (C-C motif) ligand 5 induced by a novel 8-hydroxyquinoline derivative (91b1) suppresses tumor invasiveness in esophageal carcinoma

doi: 10.3892/ijmm.2024.5435

Figure Lengend Snippet: MTS 50 ( μ g/ml) of 91b1 and CDDP for four ESCC cell lines and two non-tumor cell lines. Results were expressed with mean ± SD from triplicate. experiments.

Article Snippet: Esophageal squamous cell carcinoma (ESCC) cell lines of Japanese origin KYSE150, KYSE450, KYSE30 and KYSE510 ( ) were purchased from Leibniz-Institut DSMZ-Deutsche Sammlung von Mikroorganismen und Zellkulturen GmbH.

Techniques:

Journal: Oncology Letters

Article Title: JAK/STAT3 signaling pathway mediates endothelial-like differentiation of immature dendritic cells

doi: 10.3892/ol.2015.3752

Figure Lengend Snippet: Cell morphology of the endothelial-like differentiation of iDCs induced by KYSE450 and KYSE70 supernatant. PBMCs were incubated in rhGM-CSF/rhIL-4 and 40% supernatant of KYSE450 (highly differentiated ESCC cells) or KYSE70 (poorly differentiated ESCC cells) was added at the end of day 2. The cells of the KYSE450 and KYSE70 groups had no obvious morphological change, similar to control DCs, following 4 days of induction. The cells of the KYSE450 group exhibited a fusiform shape following 8 days of induction. The KYSE70 group of cells was arranged as strip-shaped and funicular structures, distinctly different to the control dendritic cells, following 8 days of induction with the KYSE70 supernatant (magnification, ×200). DCs; dendritic cells; PBMCs, peripheral blood mononuclear cells.

Article Snippet: The KYSE450 and KYSE70 ESCC cell lines (Nanjing KeyGEN Biotech.

Techniques: Incubation, Stripping Membranes

Journal: Oncology Letters

Article Title: JAK/STAT3 signaling pathway mediates endothelial-like differentiation of immature dendritic cells

doi: 10.3892/ol.2015.3752

Figure Lengend Snippet: Expression of endothelial cell markers vWF and CD144 increased following induction by KYSE450 and KYSE70 supernatant. (A) Immunofluoresence was used to detect the expression of vWF and CD144 in control DCs and the KYSE450 and KYSE70 groups (magnification, ×200). Data are the mean ± standard deviation (SD), n=4, ***P<0.001 vs. control DCs. (B) RT-qPCR detection of mRNA levels of vWF and CD144 in control DCs and the KYSE450 and KYSE70 groups. Data are the mean ± SD, n=3, **P<0.01, ***P<0.001 vs. control DCs. (C) Dil-Ac-LDL uptake was enhanced following induction by KYSE450 and KYSE70 supernatant (magnification, ×200). Data are the mean ± SD, n=4, ***P<0.001 vs. control DCs. DCs, dendritic cells; vWF, von Willebrand factor; CD144, cluster of differentiation 144; DiI-Ac-LDL, DiI-labeled acetylated low-density lipoprotein; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: The KYSE450 and KYSE70 ESCC cell lines (Nanjing KeyGEN Biotech.

Techniques: Expressing, Standard Deviation, Quantitative RT-PCR, Labeling, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: JAK/STAT3 signaling pathway mediates endothelial-like differentiation of immature dendritic cells

doi: 10.3892/ol.2015.3752

Figure Lengend Snippet: Activation of JAK/STAT3 signaling by KYSE 450 and KYSE70 supernatant. (A) RT-qPCR analysis revealed that JAK and STAT3 mRNA expression levels increased significantly following incubation with KYSE450 and KYSE70 supernatant. Additionally, the known targets of JAK/STAT3 signaling, VEGF and IL-6 had significantly increased mRNA expression. Data are the mean ± SD (n=3). *P<0.05, **P<0.01, ***P<0.001 vs. control dendtritic cells (DCs). (B) Immunofluorescence detection of the activation of JAK and STAT3 in control DCs and the KYSE450 and KYSE70 groups (magnification, ×200). Data are the mean ± SD (n=3). ***P<0.001 vs. control DCs. JAK, janus tyrosine kinase; VEGF, vascular endothelial growth factor; STAT3, signal transducer and activator of transcription 3.

Article Snippet: The KYSE450 and KYSE70 ESCC cell lines (Nanjing KeyGEN Biotech.

Techniques: Activation Assay, Quantitative RT-PCR, Expressing, Incubation, Immunofluorescence

Journal: Oncology Letters

Article Title: JAK/STAT3 signaling pathway mediates endothelial-like differentiation of immature dendritic cells

doi: 10.3892/ol.2015.3752

Figure Lengend Snippet: Blocking the activation of JAK and STAT3 inhibited endothelial-like differentiation of iDCs. (A) Addition of 10 µmol AG490 at the end of day 2, decreased the expression of vWF and CD144 in iDCs induced by KYSE450 and KYSE70 supernatant. RT-qPCR results shown are the mean ± SD (n=3). **P<0.01, ***P<0.001 vs. KYSE450 group. **P<0.01, ***P<0.001 vs.KYSE70 group. (B) Immunofluorescence detection of the protein expression of the EC markers CD144 and vWF in the KYSE450, KYSE450 + AG, KYSE70, and KYSE70 + AG groups (magnification, ×200). Data are the mean ± SD (n=3). ***P<0.001 vs. KYSE450 group. ***P<0.001 vs. KYSE70 group. (C) Dil-Ac-LDL uptake decreased after induction by KYSE450 + AG490 and KYSE70 + AG490. Original magnification, ×200. Results shown are the mean ± SD (n=4). ***P<0.001 vs. KYSE450 group. ***P<0.001 vs. KYSE70 group. AG, AG490; iDCs, immature dendritic cells; vWF, von Willebrand factor; CD144, cluster of differentiation 144; DiI-Ac-LDL, DiI-labeled acetylated low-density lipoprotein; RT-qPCR, reverse transcription-quantitative polymerase chain reaction.

Article Snippet: The KYSE450 and KYSE70 ESCC cell lines (Nanjing KeyGEN Biotech.

Techniques: Blocking Assay, Activation Assay, Expressing, Quantitative RT-PCR, Immunofluorescence, Labeling, Real-time Polymerase Chain Reaction

Journal: Cancer Science

Article Title: Sensitivities to various epidermal growth factor receptor‐tyrosine kinase inhibitors of uncommon epidermal growth factor receptor mutations L861Q and S768I: What is the optimal epidermal growth factor receptor‐tyrosine kinase inhibitor?

doi: 10.1111/cas.12980

Figure Lengend Snippet: Sensitivities to various EGFR‐TKI in the KYSE270 and KYSE450 cell lines. (a) Growth inhibitory curves of various EGFR‐TKI in the KYSE270 and KYSE450 cell lines. The cells were exposed to each concentration of various EGFR‐TKI for 72 h, and the growth inhibitory effects were evaluated using an MTT assay. Both the KYSE270 (L861Q) and KYSE450 (S768I) cell lines were most sensitive to afatinib and least sensitive to erlotinib. Osimertinib was as effective for the KYSE270 cell line as afatinib, but was much less effective for the KYSE450 cell line than afatinib. Lines, mean of independent triplicate experiments. (b) IC 50 values of various EGFR‐TKI in the KYSE270 and KYSE450 cell lines. The IC 50 values of erlotinib, afatinib and osimertinib in the KYSE 270 cell line (L861Q) were 0.7, 0.004 and 0.03 μM, respectively, while those of erlotinib, afatinib and osimertinib in the KYSE 450 cell line (S768I) were 1.1, 0.004 and 0.4 μM, respectively.

Article Snippet: The KYSE270 and KYSE450 cell lines (human esophageal cancer cell lines) were maintained in a 1:1 mixture of RPMI1640 and F12 (Nissui Pharmaceutical, Tokyo, Japan) with 2% FBS according to a previously reported method., , According to the Catalogue of Somatic Mutations in Cancer (COSMIC) database ( http://cancer.sanger.ac.uk/cancergenome/projects/cosmic/ ), these cell lines carry uncommon EGFR mutations (KYSE270, L861Q and KYSE450, S768I, respectively).

Techniques: Concentration Assay, MTT Assay

Journal: Cancer Science

Article Title: Sensitivities to various epidermal growth factor receptor‐tyrosine kinase inhibitors of uncommon epidermal growth factor receptor mutations L861Q and S768I: What is the optimal epidermal growth factor receptor‐tyrosine kinase inhibitor?

doi: 10.1111/cas.12980

Figure Lengend Snippet: Western blotting in the KYSE270 and KYSE450 cell lines. (a) Phosphorylation of epidermal growth factor receptor (EGFR). Three hours after the cells were treated with the indicated drug concentrations, the samples were collected. In both the KYSE270 (L861Q) and the KYSE450 (S768I) cell lines, afatinib markedly inhibited the phosphorylation of EGFR, compared with erlotinib. Osimertinib intermediately inhibited the phosphorylation. In the KYSE450 cell line, especially, the EGFR inhibitory effect of osimertinib was much weaker than that of afatinib. β‐actin was used as an internal control. p‐EGFR, phospho‐EGFR. (b) Expression of apoptosis‐related molecules. Twenty‐four hours after the cells were exposed to each drug (0.1 μM), the samples were collected. In the KYSE270 cell line (L861Q), afatinib and osimertinib induced an expression of cleaved caspase‐3, whereas only afatinib induced the expression of cleaved caspase‐3, in the KYSE450 cell line (S768I). β‐actin was used as an internal control. c‐Caspase3, cleaved caspase‐3.

Article Snippet: The KYSE270 and KYSE450 cell lines (human esophageal cancer cell lines) were maintained in a 1:1 mixture of RPMI1640 and F12 (Nissui Pharmaceutical, Tokyo, Japan) with 2% FBS according to a previously reported method., , According to the Catalogue of Somatic Mutations in Cancer (COSMIC) database ( http://cancer.sanger.ac.uk/cancergenome/projects/cosmic/ ), these cell lines carry uncommon EGFR mutations (KYSE270, L861Q and KYSE450, S768I, respectively).

Techniques: Western Blot, Phospho-proteomics, Control, Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Construction of a risk signature using the least absolute shrinkage and selection operator (LASSO) analysis. Partial likelihood deviances for (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Coefficient profiles of senescence‐related gene pairs for (C) ESCC and (D) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Selection

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Kaplan–Meier analysis of high‐ and low‐risk patients (red and blue, respectively) with (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Receiver operating characteristic curves for (C) ESCC and (D) EAC. Survival risk curves (top) and sand scatter plots (bottom) for (E) ESCC and (F) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Correlations of immune microenvironments evaluated using ESTIMATE. (A) Immune score and (B) ESTIMATE score for oesophageal squamous cell carcinoma (ESCC). (C) Immune score and (D) ESTIMATE score for oesophageal adenocarcinoma (EAC). Relationships between risk and immune scores for (E) ESCC and (H) EAC. Relationships between risk and ESTIMATE scores for (F) ESCC and (G) EAC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques:

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Random forest error rates (left graphs) and relative importance (right graphs) for (A) oesophageal squamous cell carcinoma (ESCC) and (B) oesophageal adenocarcinoma (EAC). Expression of IRF5 and BMI1 in (C) ESCC, (D) EAC and (E) EC.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Expressing

Journal: Journal of Cellular and Molecular Medicine

Article Title: Multi‐Omics Analysis of Aberrances and Functional Implications of IRF5 in Digestive Tract Tumours

doi: 10.1111/jcmm.70433

Figure Lengend Snippet: Expression analyses of IRF5 in four ESCC cell lines using western blotting (A, B). The efficiency of IRF5 ‐knockdown in KYSE150 cells was determined using western blotting (C, D). Ctrl: No siRNA infection; NC: Negative control. Statistical analyses of n = 3 independent experiments were assessed. Results are shown as mean ± SD, ns p ≥ 0.05, * p < 0.05, ** p < 0.01, *** p < 0.001.

Article Snippet: Human ESCC cell lines (KYSE150, KYSE180, KYSE410 and KYSE450) and the human embryonic oesophageal cell line (SHEE) were purchased from iCell Bioscience Inc. (Shanghai, China).

Techniques: Expressing, Western Blot, Knockdown, Infection, Negative Control