Journal: International journal of molecular sciences

Article Title: Genetic Interference of FGFR3 Impedes Invasion of Upper Tract Urothelial Carcinoma Cells by Alleviating RAS/MAPK Signal Activity.

doi: 10.3390/ijms24021776

Figure Lengend Snippet: Figure 1. Up-regulation of FGFR3 in renal pelvis tissues from Caucasian patients with urothelial carcinoma. Tissue microarray slides containing sections from Caucasian patients with renal pelvis urothelial carcinoma at stages T1–2 (n = 62), T3 (n = 17), and T4 (n = 5) were subjected to immuno- histochemistry (IHC) staining with FGFR3. (A) Representative FGFR3-positive stained images from stages T1–2, T3, and T4. (B) Quantification of FGFR3 expression levels from IHC staining results (scores were from 1 to 4). Data are shown in box plots displaying 25% to 75% percentile. * indicates significance between the indicated groups. (C) Heterogeneous distribution of FGFR3 in normal renal tubular epithelium. Note that proximal renal tubules (black arrows) usually show weak FGFR3 expression (score 1+), while distal tubules (white arrowheads) in the same specimen likely display stronger FGFR3 signals (score 2+~3+).

Article Snippet: To compare the effect of FGFR3 gene silencing with that of receptor kinase inhibition, the cells were treated with PD173074 (Cat. No. T2642, Targetmol, Wellesley Hills, MA, USA) at 10 nM.

Techniques: Microarray, Immunohistochemistry, Staining, Expressing

Journal: International journal of molecular sciences

Article Title: Genetic Interference of FGFR3 Impedes Invasion of Upper Tract Urothelial Carcinoma Cells by Alleviating RAS/MAPK Signal Activity.

doi: 10.3390/ijms24021776

Figure Lengend Snippet: Figure 2. Up-regulation of FGFR3 in renal pelvis tissues from Asian patients with upper tract urothelial carcinoma (UTUC). Renal pelvis tissues were collected from Asian patients diagnosed with UTUC at different TNM stages, including T1–2 (n = 44), T3 (n = 38), and T4 (n = 30). The formalin- fixed paraffin-embedded sections were subjected to immunohistochemistry (IHC) analysis of FGFR3 expression. (A) Representative FGFR3-positive stained images from stages T1–2, T3, and T4 and that of archived normal kidney pelvis tissues. (B) Analysis of FGFR3 IHC scoring results. (C) Statistical analysis of ethnic difference in FGFR3 expression at different UTUC stages. Data are shown in box plots displaying 25% to 75% percentiles. * indicates significance between the indicated groups.

Article Snippet: To compare the effect of FGFR3 gene silencing with that of receptor kinase inhibition, the cells were treated with PD173074 (Cat. No. T2642, Targetmol, Wellesley Hills, MA, USA) at 10 nM.

Techniques: Immunohistochemistry, Expressing, Staining

Journal: International journal of molecular sciences

Article Title: Genetic Interference of FGFR3 Impedes Invasion of Upper Tract Urothelial Carcinoma Cells by Alleviating RAS/MAPK Signal Activity.

doi: 10.3390/ijms24021776

Figure Lengend Snippet: Figure 3. Suppressive effect of FGFR3 gene silencing on proliferation of cultured UTUC cells. Two UTUC cell lines, BFTC-909 (A) and UM-UC-14 (B), were treated with FGFR3-targeted siRNA or scramble non-target control (NTC), followed by a WST-1-based proliferation assay. Optical den- sity (OD) measured is expressed as mean ± SEM (n = 3). *** p < 0.001 compared with respective NTC groups.

Article Snippet: To compare the effect of FGFR3 gene silencing with that of receptor kinase inhibition, the cells were treated with PD173074 (Cat. No. T2642, Targetmol, Wellesley Hills, MA, USA) at 10 nM.

Techniques: Cell Culture, Control, Proliferation Assay

Journal: International journal of molecular sciences

Article Title: Genetic Interference of FGFR3 Impedes Invasion of Upper Tract Urothelial Carcinoma Cells by Alleviating RAS/MAPK Signal Activity.

doi: 10.3390/ijms24021776

Figure Lengend Snippet: Figure 4. Suppressive effect of FGFR3 gene silencing on migration of cultured UTUC cells. Two UTUC cell lines, BFTC-909 (A) and UM-UC-14 (B), were treated with FGFR3-targeted siRNA or scramble non-target control (NTC), followed by consecutive microscopic documentation for the post-wounding closure at the indicated time points.

Article Snippet: To compare the effect of FGFR3 gene silencing with that of receptor kinase inhibition, the cells were treated with PD173074 (Cat. No. T2642, Targetmol, Wellesley Hills, MA, USA) at 10 nM.

Techniques: Migration, Cell Culture, Control

Journal: International journal of molecular sciences

Article Title: Genetic Interference of FGFR3 Impedes Invasion of Upper Tract Urothelial Carcinoma Cells by Alleviating RAS/MAPK Signal Activity.

doi: 10.3390/ijms24021776

Figure Lengend Snippet: Figure 7. Effects of FGFR3 gene silencing and FGFR3 kinase inhibition on apoptosis of cultured UTUC cells. (A) BFTC-909 cells were treated with either FGFR3-targeted siRNA, scramble non-target control (NTC), or PD173074 at 10 nM for 48 h and subjected to TUNEL fluorescent staining. The TUNEL-positive apoptotic cells show green fluorescence, while DAPI blue fluorescence indicates nuclei of all cells. Original magnification: 100×. (B) Morphometry analysis on TUNEL-positive rates in the treated cells. Density data are expressed as mean ± SEM (n = 8). *** indicates p < 0.001 vs. control group. ND, not detectable.

Article Snippet: To compare the effect of FGFR3 gene silencing with that of receptor kinase inhibition, the cells were treated with PD173074 (Cat. No. T2642, Targetmol, Wellesley Hills, MA, USA) at 10 nM.

Techniques: Inhibition, Cell Culture, Control, TUNEL Assay, Staining

Journal: International journal of molecular sciences

Article Title: Genetic Interference of FGFR3 Impedes Invasion of Upper Tract Urothelial Carcinoma Cells by Alleviating RAS/MAPK Signal Activity.

doi: 10.3390/ijms24021776

Figure Lengend Snippet: Figure 8. A mechanistic scheme depicting the effects of FGFR3 gene silencing on RAS/mitogen- activated protein kinase (MAPK) signaling and renal pelvis tumorigenesis. Upon stimulation by fibroblast growth factor (FGF), the intracellular tyrosine kinase domain of FGF receptor 3 (FGFR3) may deliver the mitogenic signal through RAS/MAPK cascade and downstream mediator activation, including extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK). Hence, FGFR3 overexpression and activation may enhance expression of epithelial–mesenchymal transition (EMT) transcription factors and mediators, which eventually contribute to tumor progression. Our findings suggest that FGFR3 siRNA delivery is a useful strategy for alleviating the RAS/MAPK signaling axis and EMT marker expression in renal pelvis urothelial cells, thereby suppressing their metastatic activity and tumor progression.

Article Snippet: To compare the effect of FGFR3 gene silencing with that of receptor kinase inhibition, the cells were treated with PD173074 (Cat. No. T2642, Targetmol, Wellesley Hills, MA, USA) at 10 nM.

Techniques: Activation Assay, Over Expression, Expressing, Marker, Activity Assay

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: Expression of FGF1 in CRC and paired normal tissues. (A) FGF1 mRNA levels in CRC and normal tissues in Skrzypczak’s datasets. (B) Comparison of FGF1 mRNA expression in CRC and normal tissues across 16 Oncomine datasets. (C) Representative IHC images showing in situ FGF1 expression in CRC and normal tissues (scale bar = 100μm). (D) IHC scores of FGF1 in CRC vs normal tissues. (E) OS of FGF1(+) and FGF1(-) in CRC patients in TCGA dataset searched via GEPIA platform. (F) OS of FGF1(+) and FGF1(-) in CRC patients. *** P < 0.001.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: Expressing, Comparison, In Situ

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: Statistics of FGF1 expression in 135 CRC tissues and adjacent normal tissue.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: Relationship between FGF1 and clinic-pathological factors in CRC patients.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques:

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: Results of univariate and multivariate analyses of underwent gastrectomy patients’ survival by Cox’s proportional hazard model.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: Effect of FGF1 on survival. (A) IHC images showing in situ FGF1 expression in CRC tissues (scale bar = 100μm). Negative (a), Weak (b), Moderate (c), Strong (d). (B) IHC scores of FGF1 in TNM stage I-II vs stage III-IV. (C) IHC scores of FGF1 in nLNM vs LNM. (D, E) OS of FGF1(+) and FGF1(-) CRC patients with TNM staging (D) I-II and (E) III-IV. nLNM, no lymph node metastasis; LNM, lymph node metastasis. ** P < 0.01.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: In Situ, Expressing

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: Subgroup analysis for the influence factor of survival duration of CRC patients according to FGF1 expression. * P < 0.05.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: Expressing

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: The relationship between FGF1 and p-S6K1.Nomograms to predict survival of CRC patients. (A) Correlation analysis of FGF1 and S6K1 at gene level in CRC tissues from TCGA datasets by GEPIA platform. (B) Representative IHC images showing in situ p-S6K1 expression in CRC and normal tissues (scale bar = 100μm). (C) IHC scores of p-S6K1 in CRC vs normal tissues. (D–G) Correlation between FGF1 and p-S6K1 at protein level in (D) CRC tissue, (E) normal tissue, (F) TNM stage I-II tissue and (G) TNM stage III-IV tissue. (H) Stratification of 135 pairs of CRC and normal tissues into cluster 1 and cluster 2 according to FGF1 and p-S6K1 IHC scores. *** P < 0.001.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: In Situ, Expressing

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: FGF1 regulates CRC cell growth in an mTOR-S6K1 pathway dependent manner. (A) Immunoblot showing FGF1, p-mTOR, mTOR, p-S6K1 and S6K1, β-Actin protein levels in CRC cells transfected with FGF1-shRNA. (B) Immunoblot result of FGF1/β-Actin, p-mTOR/mTOR and p-S6K1/S6K1 were semi-quantified by ImageJ. Data are presented as mean ± SD. NC, negative control; KD, FGF1-shRNA. * P < 0.05, ** P < 0.01.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: Western Blot, Transfection, shRNA, Negative Control

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: FGF1 promotes proliferation and migration ability of CRC cells. (A, B) Colony formation capacity (A) and migration rates (B) of FGF1-KD CRC cells. CRC, colorectal cancer. (C) Wound healing assays were carried out at 24h after transfection in 6-well plates. The gap width was measured using Open Lab software. (D) The wound rate was calculated and displayed graphically according to the measured results by Open Lab software. NC, negative control; KD, FGF1-shRNA. Data are presented as mean ± SD (n=3). Ns, no significance, * P < 0.05.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: Migration, Transfection, Software, Negative Control, shRNA

Journal: Frontiers in Oncology

Article Title: The Association of Aberrant Expression of FGF1 and mTOR-S6K1 in Colorectal Cancer

doi: 10.3389/fonc.2021.706838

Figure Lengend Snippet: FGF1 induces CRC tumor growth in vivo . (A, B) Total body weight (A) and tumor volume (B) of the mice during the experiment. (C) Representative pictures of subcutaneous tumors harvested from NC and FGF1-KD group. (D) The weights of tumor masses. (E) Net body weight after subtracting the respective tumor weights. (F) Relative FGF1 mRNA levels in the tumors and the tumor weight correlation. (G) Stratification of mice into cluster 1 (blue) and cluster 2 (grey) according to tumor weight and FGF mRNA levels. (H) Percentage of NC and FGF1-KD mice in each cluster. Data are presented as mean ± SD (n=5). CRC, colorectal cancer. NC, negative control; KD, FGF1-shRNA. * P < 0.05.

Article Snippet: After blocking with 5% non-fat milk for 1h, the membranes were probed overnight with FGF1 (1:1000, BOSTER, China), p-mTOR (1:1000, Cell Signaling Technology, USA), mTOR (1:1000, Bioss, China), p-S6K1 (1:1000, Cell Signaling Technology, USA), and S6K1 (1:1000, Bioss, China) and β-Actin (1:5000, BOSTER, China) antibodies at 4°C with gentle shaking, followed by horseradish peroxidase-conjugated secondary antibodies.

Techniques: In Vivo, Negative Control, shRNA