Journal: Cancer Cell International

Article Title: Silenced lncRNA SNHG14 restrains the biological behaviors of bladder cancer cells via regulating microRNA-211-3p/ESM1 axis

doi: 10.1186/s12935-020-01717-7

Figure Lengend Snippet: PCR primer sequence

Article Snippet: The membrane was blocked with 5% skim milk powder, incubated with primary antibodies ESM1 (1: 200, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) and GAPDH (1: 1000, Cell Signaling Technology, Beverly, MA, USA), and with the secondary antibody, horseradish peroxidase-labeled IgG (1: 1000, Wuhan Boster Biological Technology Co., Ltd., Hubei, China).

Techniques: Sequencing

Journal: Cancer Cell International

Article Title: Silenced lncRNA SNHG14 restrains the biological behaviors of bladder cancer cells via regulating microRNA-211-3p/ESM1 axis

doi: 10.1186/s12935-020-01717-7

Figure Lengend Snippet: Elevated SNHG14, ESM1 and decreased miR-211-3p are found in BCa tissues. a The expression of SNHG14 in bladder tissues of the two groups detected via RT-qPCR; b The expression of miR-211-3p in bladder tissues of the two groups detected via RT-qPCR; c The expression of ESM1 mRNA in bladder tissues of the two groups detected via RT-qPCR; d Protein bands of ESM1 in bladder tissues of the two groups detected via western blot analysis; e The expression of ESM1 protein in bladder tissues of the two groups detected via western blot analysis. f The effect of SNHG14 expression on the prognosis of BCa patients; g The effect of miR-211-3p expression on the prognosis of BCa patients; h The effect of ESM1 expression on the prognosis of BCa patients; n = 62; * vs the normal group, P < 0.05; The measurement data were expressed as mean ± standard deviation. The data were compared via using t test in two groups

Article Snippet: The membrane was blocked with 5% skim milk powder, incubated with primary antibodies ESM1 (1: 200, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) and GAPDH (1: 1000, Cell Signaling Technology, Beverly, MA, USA), and with the secondary antibody, horseradish peroxidase-labeled IgG (1: 1000, Wuhan Boster Biological Technology Co., Ltd., Hubei, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation

Journal: Cancer Cell International

Article Title: Silenced lncRNA SNHG14 restrains the biological behaviors of bladder cancer cells via regulating microRNA-211-3p/ESM1 axis

doi: 10.1186/s12935-020-01717-7

Figure Lengend Snippet: The expression of ESM1 in BCa is related to the patients’ TNM stage, tumor invasion stage and lymph node metastasis

Article Snippet: The membrane was blocked with 5% skim milk powder, incubated with primary antibodies ESM1 (1: 200, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) and GAPDH (1: 1000, Cell Signaling Technology, Beverly, MA, USA), and with the secondary antibody, horseradish peroxidase-labeled IgG (1: 1000, Wuhan Boster Biological Technology Co., Ltd., Hubei, China).

Techniques: Expressing

Journal: Cancer Cell International

Article Title: Silenced lncRNA SNHG14 restrains the biological behaviors of bladder cancer cells via regulating microRNA-211-3p/ESM1 axis

doi: 10.1186/s12935-020-01717-7

Figure Lengend Snippet: SNHG14 and ESM1 are up-regulated while miR-211-3p is down-regulated in BCa cell lines. a The expression of SNHG14, miR-211-3p and ESM1 in T24, 5637, UMUC-3, EJ and SV-HVC-1 cell lines tested via RT-qPCR; b Protein bands of ESM1 in T24, 5637, UMUC-3, EJ and SV-HVC-1 cell lines tested via western blot analysis; c The expression of ESM1 protein in T24, 5637, UMUC-3, EJ and SV-HVC-1 cell lines tested via western blot analysis. * vs the SV-HVC-1 cell lines, P < 0.05. The measurement data were expressed as mean ± standard deviation. One-way ANOVA was functioned for comparison among multiple groups, and Tukey’s post hoc test was used for pairwise comparison

Article Snippet: The membrane was blocked with 5% skim milk powder, incubated with primary antibodies ESM1 (1: 200, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) and GAPDH (1: 1000, Cell Signaling Technology, Beverly, MA, USA), and with the secondary antibody, horseradish peroxidase-labeled IgG (1: 1000, Wuhan Boster Biological Technology Co., Ltd., Hubei, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Comparison

Journal: Cancer Cell International

Article Title: Silenced lncRNA SNHG14 restrains the biological behaviors of bladder cancer cells via regulating microRNA-211-3p/ESM1 axis

doi: 10.1186/s12935-020-01717-7

Figure Lengend Snippet: There is a binding relationship between SNHG14 and miR-211-3p and a targeting relationship between miR-211-3p and ESM1. a The binding sites of SNHG14 and miR-211-3p predicted via Diana tools; b The binding relationship between SNHG14 and miR-211-3p in T24 cells verified via dual luciferase reporter gene assay; c The binding relationship between SNHG14 and miR-211-3p in UMUC-3 cells verified via dual luciferase reporter gene assay. d The binding relationship between SNHG14 and miR-211-3p verified via RNA pull down assay. e The binding site of miR-211-3p and ESM1 predicted by Jefferson; f The binding relationship between miR-211-3p and ESM1 in T24 cells verified via dual luciferase reporter gene assay; g The binding relationship between miR-211-3p and ESM1 in UMUC-3 cells verified via dual luciferase reporter gene assay; * vs the mimic NC group, P < 0.05; ^ vs the Bio-NC group, P < 0.05; The measurement data were expressed as mean ± standard deviation. The data were compared via using t test in two groups. One-way ANOVA was functioned for comparison among multiple groups, and Tukey’s post hoc test was used for pairwise comparison

Article Snippet: The membrane was blocked with 5% skim milk powder, incubated with primary antibodies ESM1 (1: 200, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) and GAPDH (1: 1000, Cell Signaling Technology, Beverly, MA, USA), and with the secondary antibody, horseradish peroxidase-labeled IgG (1: 1000, Wuhan Boster Biological Technology Co., Ltd., Hubei, China).

Techniques: Binding Assay, Luciferase, Reporter Gene Assay, Pull Down Assay, Standard Deviation, Comparison

Journal: Cancer Cell International

Article Title: Silenced lncRNA SNHG14 restrains the biological behaviors of bladder cancer cells via regulating microRNA-211-3p/ESM1 axis

doi: 10.1186/s12935-020-01717-7

Figure Lengend Snippet: Silenced SNHG14 increases miR-211-3p expression and decreases ESM1 expression; elevated miR-211-3p decreases ESM1 expression in BCa cells. a The efficiency of SNHG14 and miR-211-3p in T24 cells was verified by RT-qPCR; b ESM1 mRNA expression in T24 cells detected via RT-qPCR; c Protein bands of ESM1 in T24 cells tested via western blot analysis; d The expression of ESM1 protein in T24 cells detected via western blot analysis; e The efficiency of SNHG14 and miR-211-3p in UMUC-3 cells was verified by RT-qPCR; f ESM1 mRNA expression in UMUC-3 cells detected via RT-qPCR; g Protein bands of ESM1 in UMUC-3 cells tested via western blot analysis; h The expression of ESM1 protein in UMUC-3 cells detected via western blot analysis. * vs the si-NC group, P < 0.05; ^ vs the mimic NC group, P < 0.05; and vs the si-SNHG14 + inhibitor NC group, P < 0.05; a vs the oe-NC group, P < 0.05; b vs the inhibitor NC group, P < 0.05; c vs the oe-SNHG14 + mimic NC group, P < 0.05. The experiment was repeated independently for 3 times, and the measurement data were expressed as mean ± standard deviation. One-way ANOVA was functioned for comparison among multiple groups, and Tukey’s post hoc test was used for pairwise comparison

Article Snippet: The membrane was blocked with 5% skim milk powder, incubated with primary antibodies ESM1 (1: 200, Santa Cruz Biotechnology, Inc, Santa Cruz, CA, USA) and GAPDH (1: 1000, Cell Signaling Technology, Beverly, MA, USA), and with the secondary antibody, horseradish peroxidase-labeled IgG (1: 1000, Wuhan Boster Biological Technology Co., Ltd., Hubei, China).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Standard Deviation, Comparison

Journal: Carcinogenesis

Article Title: FGF2 and EGF induce epithelial-mesenchymal transition in malignant pleural mesothelioma cells via a MAPKinase/MMP1 signal.

doi: 10.1093/carcin/bgy018

Figure Lengend Snippet: Figure 5. Pathway dependency of the sarcomatoid phenotype. Scatter scores were determined in M38K cells that (A) were treated with the Ets inhibitor YK4-279 (YK), the pan-MMP inhibitor GM6001 (GM) or vehicle (V) in combination with FGF2 and (B) were transfected with siRNAs targeting ESM1, MMP1 or control siRNA (scr) on the day before FGF2 treatment. (C) Representative micrographs of M38K cells treated with GM6001 and FGF2. (D) M38K cells were seeded into collagen-coated transwell chambers, treated with GM6001 (GM) in combination with FGF2 and allowed to invade through the collagen and the porous membrane for 72 h. Morphology (left) of (E) VMC58 cells and (F) Meso84 cells treated with the FGFR inhibitor nintedanib (Nin). Scatter scores (right) of the same cell lines treated with the FGFR inhibitors nin- tedanib, PD-166866 (PD), ponatinib (Pon) or BGJ-398 (BGJ), the EGFR inhibitor erlotinib (Erlo), the MEK inhibitors selumetinib (Sel) and UO-126 (UO), the AKT inhibitor MK-2206 (MK), the PI3K inhibitor LY-294002 (LY), the NFκB inhibitor BAY11-7082 (BAY), the FAK inhibitor BI-853520 (BI) or DMSO as control (Co) for 48 h. VMC58 (G) and Meso84 (H) cells were treated with nintedanib (Nin) or PD-166866 (PD) for 48 h, and expression of the indicated genes was analyzed by qPCR in comparison with vehicle- treated controls. Data were normalized to GAPDH. *P < 0.05, **P < 0.01, ***P < 0.001, treated versus control, one-way ANOVA with Dunnett’s post-test. Scale bar = 100 µm.

Article Snippet: SiRNAs (10 nM, scrambled (D001810-15-05, GE Dharmacon), MMP1 (SC41552), ESM1 (SC-40543), Santa Cruz Biotechnology, Dallas, TX) and Lipofectamine (RNAiMAX, Thermo Fisher Scientific) were mixed with serum-free RPMI medium in six-well plates and incubated for 30 min before 5-fold volumes of medium (with 10% FBS) containing 1 × 105 cells per well were added.

Techniques: Transfection, Control, Membrane, Expressing, Comparison