Journal: Frontiers in Oncology

Article Title: FGF19 Is Coamplified With CCND1 to Promote Proliferation in Lung Squamous Cell Carcinoma and Their Combined Inhibition Shows Improved Efficacy

doi: 10.3389/fonc.2022.846744

Figure Lengend Snippet: FGF19 enhances CCND1 expression by FGF19-FGFR4-ERK1/2 axis in LUSC cells. (A) Recombinant human FGF19 (rhFGF19) (25 ng/ml) promoted expression of CCND1 in SK-MES-1 and H520 cells (serum starved for 12 h before treatment) in a time-dependent manner. (B) CCND1 mRNA expression levels in LUSC cell lines after treatment with rhFGF2 for 12 h. (C) Expression of CCND1 in FGF19 overexpression LUSC cell line. (D) Expression of CCND1 in FGF19 knockdown LUSC cell line. (E) A panel of inhibitors against a number of signaling pathways was used to dissect the leading factors of regulated by FGF19. (F) Western blot and (G) qPCR analysis of CCND1 expression in H520 and HCC95 LUSC cells after treatment of FGF19, FGF19 and SCH772984, FGF19 & BLU9931, or DMSO as control. (H) Effects of the FGF19/FGFR4 pathway on protein levels of CCND1, p-FGFR4, and p-ERK1/2 by western blot analysis. H520, SK-MES-1 and HCC95 cells were treated with FGF19 (25 ng/ml, 0/0.5/6/12 h) to activate the FGF19/FGFR4 signaling pathway. Data were shown as mean ± SD bars and compared by unpaired t -test. ** p < 0.01; **** p < 0.0001; ns, not significant.

Article Snippet: Antibodies for western blot were as follows: β-Actin (Proteintech,66009-1-Ig), β-Tubulin ( Proteintech, 66240-1-Ig ) , FGF19 (R&D System, AF969), FGFR4 (R&D System, MAB6852), FRS2 (Abcam, ab10425), pFRS2 (R&D System, AF5126), goat anti-mouse IgG-HRP (Jackson ImmunoResearch, 115-035-003), goat anti-rabbit IgG-HRP (Jackson ImmunoResearch, 111-035-003), donkey anti-Goat IgG-HRP (Sangon Biotech, D110115) and other antibodies were obtained from the Cell Signaling Technology.

Techniques: Expressing, Recombinant, Over Expression, Knockdown, Protein-Protein interactions, Western Blot, Control

Journal: Frontiers in Oncology

Article Title: FGF19 Is Coamplified With CCND1 to Promote Proliferation in Lung Squamous Cell Carcinoma and Their Combined Inhibition Shows Improved Efficacy

doi: 10.3389/fonc.2022.846744

Figure Lengend Snippet: The schematic model of this study: underlying mechanism that link neighboring oncogene FGF19 & CCND1 co-amplification in the development of LUSC. FGF19 was co-amplified and co-expressed with its neighboring gene CCND1 in a subset of LUSC. Overexpression of FGF19 activates FGFR4 and leads to phosphorylation of FRS2 and ERK1/2, which further strengthens the increase in CCND1 expression caused by amplification. These lead to the phosphorylation of RB by CCND1-CDK4/6 complex, causing unrestrained transcription factor E2F, which promotes cell cycle progression and lead to the malignant proliferation of LUSC. While co-targeting FGFR4 and CDK4/6 by BLU9931 and palbociclib potentiated the growth inhibition and arrested cells in G1 phase.

Article Snippet: Antibodies for western blot were as follows: β-Actin (Proteintech,66009-1-Ig), β-Tubulin ( Proteintech, 66240-1-Ig ) , FGF19 (R&D System, AF969), FGFR4 (R&D System, MAB6852), FRS2 (Abcam, ab10425), pFRS2 (R&D System, AF5126), goat anti-mouse IgG-HRP (Jackson ImmunoResearch, 115-035-003), goat anti-rabbit IgG-HRP (Jackson ImmunoResearch, 111-035-003), donkey anti-Goat IgG-HRP (Sangon Biotech, D110115) and other antibodies were obtained from the Cell Signaling Technology.

Techniques: Amplification, Over Expression, Phospho-proteomics, Expressing, Inhibition