Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: Top 20 upregulated protein molecules modulated by CDDO-Me in Ec109 cells

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: The top ten canonical signaling pathways regulated by CDDO-Me in Ec109 cells analyzed by ingenuity pathway analysis. Abbreviations: CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid; eIF2, eukaryotic initiation factor 2; p70S6K, p70S6 kinase; mTOR, mammalian target of rapamycin; RAN, ras-related nuclear protein; Nrf2, nuclear factor (erythroid-derived 2)-like 2.

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques: Protein-Protein interactions, Derivative Assay

Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: Representative blots of protein levels of VKORC1, CaMKIIα, NPLOC4, PSME3, and Dynamin 2 in various human ESCC cell lines and normal human esophageal epithelial cell line (Het-1A) were determined using Western blot analysis. β-Actin served as loading controls. Abbreviations: VKORC1, vitamin K epoxide reductase complex subunit 1; CaMKIIα, calcium/calmodulin-dependent protein kinase type II subunit alapha; NPLOC4, nuclear protein localization protein 4 homolog; PSME3, proteasome activator complex subunit 3; ESCC, esophageal squamous cell carcinoma.

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques: Western Blot

Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: Mitochondrial dysfunction signaling pathway regulated by CDDO-Me in Ec109 cells. Notes: Ec109 cells were treated with 0.5 μM CDDO-Me for 24 hours and the protein samples were subject to quantitative proteomic analysis. Red indicates an upregulation; green indicates a downregulation. The intensity of green and red molecule colors indicates the degree of down- or upregulation, respectively. Solid arrows indicate direct interaction and dashed arrows indicate indirect interaction. Abbreviations: CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid.

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: mTOR signaling pathway regulated by CDDO-Me in Ec109 cells. Notes: Ec109 cells were treated with 0.5 μM CDDO-Me for 24 hours and the protein samples were subject to quantitative proteomic analysis. Red indicates an upregulation; green indicates a downregulation. The intensity of green and red molecule colors indicates the degree of down- or upregulation, respectively. Solid arrows indicate direct interaction and dashed arrows indicate indirect interaction. Abbreviation: CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid; mTOR, mammalian target of rapamycin.

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques:

Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: The effect of knockdown of CaMKIIα on the CDDO-Me induced apoptosis in human ESCC cells. A. Percentages of specific cell populations showed in dot plots and apoptotic cells showed in bar graphs for the silencing of CaMKIIα in Ec109 and KYSE30 cells with or without 0.5 μM CDDO-Me treatment for 24 hours. B. Representative blots of bcl-2, bax, and cleaved caspase-3 for the proteinlysate samples, which were prepared from Ec109 and KYSE30 cells treated with CaMKIIα siRNAs or 0.5 μM CDDO-Me for 24 hours. Abbreviation: CaMKIIα, calcium/calmodulin-dependent protein kinase type II subunit alapha; CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid; ESCC, esophageal squamous cell carcinoma; bcl-2, B-cell lymphoma-2; bax, bcl-2 associated X protein; siRNA , small interfering RNA.

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques: Knockdown, Small Interfering RNA

Journal: American Journal of Translational Research

Article Title: The therapeutic response of CDDO-Me in the esophageal squamous cell carcinoma (ESCC) cells is mediated by CaMKIIα

doi:

Figure Lengend Snippet: The effect of knockdown of CaMKIIα on the CDDO-Me induced autophagy in human ESCC cells. A. Percentages of specific cell populations showed in dot plots and autophagic cells showed in bar graphs for the silencing of CaMKIIα in Ec109 and KYSE30 cells with or without 0.5 μM CDDO-Me treatment for 24 hours. B. CDDO-Me-induced autophagic death in the silencing of CaMKIIα in Ec109 and KYSE30 cellsdetermined by confocal microscopy. The level of autophagy was evaluated using a lysosome-specific fluorescence dye. The confocal microscopic images of autophagic Ec109 and KYSE 30 cells (stained in green) are also shown. C. Representative blots of beclin-1 and LC3I/II for the proteinlysate samples, which were prepared from Ec109 and KYSE30 cells treated with CaMKIIα siRNAs or 0.5 μM CDDO-Me for 24 hours. Abbreviation: CaMKIIα, calcium/calmodulin-dependent protein kinase type II subunit alapha; CDDO-Me, methyl ester of 2-cyano-3,12-dioxoolean-1,9-dien-28-oic acid; ESCC, esophageal squamous cell carcinoma; siRNA, small interfering RNA; LC 3, microtubule-associated protein 1A/1B-light chain 3.

Article Snippet: Human ESCC Ec109 cells was obtained from AddexBio Inc. (San Diego, CA, USA), KYSE70 and KYSE30 cells were obtained from Sigma-Aldrich Co (St Louis, MO, USA).

Techniques: Knockdown, Confocal Microscopy, Fluorescence, Staining, Small Interfering RNA

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

Journal: Nature Communications

Article Title: Nuclear ANLN regulates transcription initiation related Pol II clustering and target gene expression

doi: 10.1038/s41467-025-56645-9

Figure Lengend Snippet: A KYSE150 cells were treated with 0.1 μM THZ1 for 3 h, ANLN (pink) and Pol II (green) clusters were observed with super-resolution imaging. Scale bar, 3 μm. B , C Effect of THZ1 on the number of different types of clusters. n = 42 (Control); n = 45 (THZ1). The error bars indicate the mean ± SDs. Two-tailed Student’s t tests were performed to determine the significance. D Relative fluorescence intensity of protein signals per cell per focal plane. n = 42 (Control); n = 45 (THZ1). The error bars indicate the mean ± SDs. Two-tailed Student’s t tests were performed to determine the significance. E , F Heatmap ( E ) and box plots ( F ) showing expression changes of all protein-coding genes (Total, n = 11376), super-enhancer-associated genes (SE, n = 652), and target genes (TG, n = 84) with or without THZ1 treatment. The error bars indicate the mean ± SDs. Two-tailed Student’s t tests were performed to determine the significance. G KYSE150 cells were treated with 0.1 μM THZ1, the mRNA expression of the indicated target genes was measured. The error bars indicate the mean ± SDs. Two-tailed Student’s t tests were performed to determine the significance. H The indicated cell lines were treated with different concentrations of THZ1 for 48 h, and cell viability was subsequently analyzed by MTS assay. The error bars indicate the mean ± SDs. I ESCC cell lines were treated with low toxicity THZ1 (0.04 μM) or/and F806 (20 μM) for 48 h, and cell viability was measured by MTS assay. n = 3 for each group. The error bars indicate the mean ± SDs. The data are representative of at least three independent experiments with similar results. Source data are provided as a Source Data file.

Article Snippet: ESCC cell lines were established and provided by Dr. Shimada Yutaka (Faculty of Medicine, Kyoto University, Japan) and Dr. Nishihira (Tohoku University School of Medicine, Japan) – .

Techniques: Imaging, Control, Two Tailed Test, Fluorescence, Expressing, MTS Assay

Journal: Cancer Medicine

Article Title: Aberrant DNA hypermethylation reduces the expression of the desmosome-related molecule periplakin in esophageal squamous cell carcinoma

doi: 10.1002/cam4.369

Figure Lengend Snippet: PPL expression was silenced by DNA methylation in ESCC. (A) Typical images of formalin-fixed, paraffin-embedded samples of ESCC, and adjacent noncancerous mucosa (normal) stained with H&E or anti-PPL antibody. (B) PPL transcript levels in paired samples from 13 ESCC samples were determined by RT-PCR. Data indicate expression relative to the mean levels of normal tissues. (C) DNA methylation of PPL as determined by pyrosequencing of paired samples from 17 patients with ESCC. (D) Expression of PPL in ESCC relative to that in normal tissue plotted against the change in DNA methylation (ratio of tumor to normal tissue) in each paired sample. (E) PPL mRNA induction in ESCC cell lines after treatment with 1 or 5 μ mol/L of 5-aza-dC. The fold increase in induction in the treated cells relative to that in the untreated cells is shown for each cell line. Data are shown as mean + SD of duplicated assays. *Difference from untreated cells was statistically significant ( P < 0.05). (F) KYSE270 cells were treated with the indicated concentrations of 5-aza-dC. The levels of methylation of PPL DNA (left Y axis, solid lines) and mRNA (right Y axis, dotted lines) are shown. Data are shown as mean ± SD of triplicated assays. *Difference from untreated cells was statistically significant ( P < 0.05). (G) KYSE270 cells were treated with 5 μ mol/L of 5-aza-dC, and stained with anti-PPL antibody (green) and DAPI for nuclear staining (red). ESCC, esophageal squamous cell carcinoma; PPL, periplakin.

Article Snippet: Human ESCC lines KYSE70, KYSE140, KYSE150, KYSE270, KYSE410, and KYSE510 were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) on 13 August 2013.

Techniques: Expressing, DNA Methylation Assay, Formalin-fixed Paraffin-Embedded, Staining, Reverse Transcription Polymerase Chain Reaction, Methylation

Journal: Cancer Medicine

Article Title: Aberrant DNA hypermethylation reduces the expression of the desmosome-related molecule periplakin in esophageal squamous cell carcinoma

doi: 10.1002/cam4.369

Figure Lengend Snippet: Forced PPL expression induced desmosome-like structures. (A) TEM images were obtained from normal human esophageal mucosa (normal) and mock-transfected or PPL-transfected KTSE270 cells at 10,000× or 30,000× magnification. Desmosomes were frequently found in normal tissues and PPL-transfected cells (arrows). In the images of the mock-transfected cells taken at 10,000× magnification, the arrow indicates adhesion plaque-like structures, which was not identified as a desmosome at higher magnification. (B) Expression of envoplakin (EVPL) in mock- or PPL- transfected KYSE270 cells. EVPL mRNA levels were shown as fold expression of the levels to normal esophageal mucosa (average of 13 mucosa = 1). Data are shown as mean + SD of three assays. (C) Expression of envoplakin (EVPL) in ESCC tissues with paired normal mucosa. PPL, periplakin; ESCC, esophageal squamous cell carcinoma.

Article Snippet: Human ESCC lines KYSE70, KYSE140, KYSE150, KYSE270, KYSE410, and KYSE510 were obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan) on 13 August 2013.

Techniques: Expressing, Transfection

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: The expression of miR-125b in ESCC tissues and cells. (A) The expression of miR-125b in ESCC and paired normal tissues was examined by qRT-PCR. (B) The expression of miR-125b in ESCC cell lines and a human esophageal epithelial cell line was examined by qRT-PCR. *P<0.05 vs. the control. ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Expressing, Quantitative RT-PCR, Control

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: The association between the expression level of miR-125b with the clinical characteristics of ESCC patients.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Expressing

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: miR-125b inhibits ESCC cell proliferation. (A) EC109 and EC9706 cells transfected with miR-125b mimics or miR-125b inhibitors significantly increased or decreased, respectively, the expression of miR-125b compared with the control. (B) EC109 and EC9706 cells transfected with miR-125b mimics or miR-125b inhibitors both exhibited a significant decrease or increase, respectively, in the proliferation rate compared with the control. *P<0.05 vs. the control. ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Transfection, Expressing, Control

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: miR-125b inhibits the ESCC cell cycle. EC109 and EC9706 cells transfected with miR-125b mimics significantly increased the G1 phase of the cell cycle compared with the control. *P<0.05 vs. the control. ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Transfection, Control

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: miR-125b induces ESCC cell apoptosis. (A) EC109 and EC9706 cells transfected with miR-125b mimics significantly increased cell apoptosis compared with the control. (B) The expression of caspase-3, Bcl-2, Bax and p27 were analyzed in ESCC cell lines. *P<0.05 vs. the control. ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Transfection, Control, Expressing

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: BMF is a direct target of miR-125b in ESCC cancer cells. (A) The prediction of the binding between miR-125b and BMF as determined using TargetScan. (B) A dual-luciferase reporter assay was performed to verify the binding of miR-125b with BMF. (C) qRT-PCR assay was performed to detect the mRNA level of BMF in EC109 and EC9706 cells treated with miR-125b mimics and miR-125b inhibitors. (D) The expression of BMF was assessed in the tumor sections. *P<0.05 vs. the control. BMF, BCL-2-modifying factor; ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Binding Assay, Luciferase, Reporter Assay, Quantitative RT-PCR, Expressing, Control

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: BMF inhibits ESCC cell proliferation. (A) A qRT-PCR assay was conducted to assess the mRNA expression of BMF. (B) Western blot analysis was performed to assess the protein expression of BMF. (C) A CCK-8 assay was used to reveal the proliferation rate in ESCC cells with si-BMF transfection. (D) The cell cycle was examined in ESCC cell lines. *P<0.05 vs. the control. BMF, BCL-2-modifying factor; ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Quantitative RT-PCR, Expressing, Western Blot, CCK-8 Assay, Transfection, Control

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: BMF induces ESCC cell apoptosis. (A) Cell apoptosis was assayed in ESCC cell lines. (B) The protein level was assayed by western blotting in ESCC cell lines *P<0.05 vs. the control. BMF, BCL-2-modifying factor; ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Western Blot, Control

Journal: Oncology Reports

Article Title: MicroRNA-125b inhibits cell proliferation and induces cell apoptosis in esophageal squamous cell carcinoma by targeting BMF

doi: 10.3892/or.2018.6413

Figure Lengend Snippet: Relationship between miR-125b and BMF in ESCC. (A) The mRNA expression of BMF in ESCC tissues compared to normal tissues. (B) The mRNA expression of BMF in ESCC cell lines (EC109 and EC9706 cells) compared to an esophageal epithelial cell line (HET-1A). (C) The protein expression of BMF in ESCC tissues compared to normal tissues. (D) The protein expression of BMF in ESCC cells (EC109 and EC9706 cells) compared to an esophageal epithelial cell line (HET-1A). (E) Data analysis of relationship between the expression of miR-125b and BMF in ESCC tissues. *P<0.05 vs. the control. BMF, BCL-2-modifying factor; ESCC, esophageal squamous cell carcinoma.

Article Snippet: In the present study, the human ESCC cell lines (EC109 and EC9706) and human esophageal epithelial cells (HET-1A) were obtained from Riken BioResource Center (Tsukuba, Japan).

Techniques: Expressing, Control

Journal: Scientific Reports

Article Title: The expression and role of TRPV2 in esophageal squamous cell carcinoma

doi: 10.1038/s41598-019-52227-0

Figure Lengend Snippet: TRPV2 controls the proliferation and cell cycle progression of ESCC cells, ( A ) TRPV2 mRNA expression was analyzed in 7 ESCC cell lines. Quantitative RT-PCR showed that TRPV2 was strongly expressed in TE15, and more strongly expressed in KYSE170 than in the other ESCC cell lines. ( B ) TRPV2 protein expression of ESCC cell lines was evaluated by western blotting. Upper panel showed full-length gels of TRPV2, and lower panel indicated the cropped image of same gels for evaluating β-actin expression (full-length image: Supplementary Fig. ). TRPV2 expression in TE15 and KYSE170 was similar in Western blotting. ( C ) TRPV2 expression of ESCC cell lines transfected with control and TRPV2 siRNA was evaluated by quantitative RT-PCR and western blotting. The blotting figure showed the cropped image of TRPV2 or β-Actin bands on same gels (full-length image: Supplementary Fig. ). TRPV2 siRNA effectively reduced TRPV2 mRNA and protein levels in TE15 and KYSE170 cells. Mean ± SEM. n = 3. * p < 0.05 (significantly different from control siRNA). ( D ) The down-regulation of TRPV2 inhibited the proliferation of TE15 and KYSE170 cells. The number of cells was counted 48 and 72 h after siRNA transfection. Mean ± SEM. n = 4. * p < 0.05 (significantly different from control siRNA). ( E ) The down-regulation of TRPV2 partially reduced cell cycle progression from the G1 to S phase in TE15 and KYSE170 cells. The cell population of sub-G1 increased in TPRV2-depleted TE15 and KYSE170 cells. Cells transfected with control or NIS siRNA were stained with propidium iodide (PI) and analyzed by flow cytometry. Mean ± SEM. n = 3. * p < 0.05 (significantly different from control siRNA).

Article Snippet: The human ESCC cell lineLYSE150 and KYSE170 was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Transfection, Control, Staining, Flow Cytometry

Journal: Scientific Reports

Article Title: The expression and role of TRPV2 in esophageal squamous cell carcinoma

doi: 10.1038/s41598-019-52227-0

Figure Lengend Snippet: TRPV2 controls the survival, migration, and invasion of GC cells. ( A ) The down-regulation of TRPV2 induced early and late apoptosis in TE15 and KYSE170 cells. Apoptosis was assessed by flow cytometry using PI/annexin V double staining. Mean ± SEM. n = 3. * p < 0.05 (significantly different from control siRNA). ( B ) Apoptosis marker expression, caspase 3 and cleaved caspase 3, of ESCC cell lines transfected with control and TRPV2 siRNA was evaluated by western blotting. The western blotting figure indicated the clopped image of the bands on same gels (full-length image: Supplementary Fig. ). The findings revealed that the expression of cleaved caspase 3, an apoptosis marker, was increased in TRPV2-depleted TE15 and KYSE170 cells. ( C ) The down-regulation of TRPV2 inhibited the migration and invasion of TE15 and KYSE170 cells. Cell migration and invasion were examined using the Boyden chamber assay. Mean ± SEM. n = 3. * p < 0.05 (significantly different from control siRNA).

Article Snippet: The human ESCC cell lineLYSE150 and KYSE170 was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Migration, Flow Cytometry, Double Staining, Control, Marker, Expressing, Transfection, Western Blot, Boyden Chamber Assay

Journal: Scientific Reports

Article Title: The expression and role of TRPV2 in esophageal squamous cell carcinoma

doi: 10.1038/s41598-019-52227-0

Figure Lengend Snippet: Signal pathways regulated by TRPV2 in ESCC cells. ( A ) ( A ) The signaling map, which was generated using an Ingenuity Pathway Analysis (IPA) software (Ingenuity Systems, Qiagen, Redwood City, CA), of “WNT/β-Catenin”, the top-ranking canonical pathway related to TRPV2 depletion according to an IPA analysis. Red and green indicate genes with expression levels that were higher or lower, respectively, than reference RNA levels. ( B ) The signaling map, which was generated by IPA software, of “Basal cell carcinoma signaling” ( C ) Verification of gene expression by real-time quantitative RT-PCR. The expression levels of four selected “WNT/TGF-βCatenin” and “Basal cell carcinoma signaling”-related genes (WNT10A, TGF-β2, TGF-β2R, and GLI) in TRPV2-depleted TE15 and KYSE170 were compared to those in control siRNA-transfected cells using real-time quantitative RT-PCR. Mean ± SEM. n = 3. * p < 0.05 (significantly different from control siRNA).

Article Snippet: The human ESCC cell lineLYSE150 and KYSE170 was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Generated, Software, Expressing, Gene Expression, Quantitative RT-PCR, Control, Transfection

Journal: Scientific Reports

Article Title: The expression and role of TRPV2 in esophageal squamous cell carcinoma

doi: 10.1038/s41598-019-52227-0

Figure Lengend Snippet: TRPV2 protein expression in human ESCCs. ( A ) Immunohistochemical staining of primary ESCC samples showed that nuclear TRPV2 expression was detected in the middle layer of the non-cancerous stratified squamous epithelium. Magnification: ×400. Bar, 100 μm. ( B ) Immunohistochemical staining of primary human ESCC samples without TRPV2 expression. Magnification: ×400. Bar, 100 μm. ( C ) Immunohistochemical staining of primary human ESCC samples with the weak cytoplasmic expression of TRPV2. Magnification: ×400. Bar, 100 μm. ( D ) Immunohistochemical staining of primary human ESCC samples with the strong cytoplasmic expression of TRPV2. Magnification: ×400. Bar, 100 μm. ( E ) Survival curve of patients after curative resection for ESCC according to the expression of TRPV2. All patients were classified into two groups according to the proportion of strong TRPV2 expression in ESCC tumors: low group: <20%, high group: ≥20%: the low group (n = 22) and high group (n = 40) in the tumor. *p < 0.05: Log-rank test. ( F ) Survival curve of ESCC patients after post-operative recurrence (n = 26) according to the expression of TRPV2. All patients were classified into two groups according to the proportion of strong TRPV2 expression in ESCC tumors: low group: <20%, high group: ≥20%: the low group (n = 8) and high group (n = 18) in the tumor. *p < 0.05: Log-rank test.

Article Snippet: The human ESCC cell lineLYSE150 and KYSE170 was obtained from the Japanese Collection of Research Bioresources Cell Bank (Osaka, Japan).

Techniques: Expressing, Immunohistochemical staining, Staining