Review

fgf19 shrna (Santa Cruz Biotechnology)

Santa Cruz Biotechnology is a verified supplier

About

News

Press Release

Team

Advisors

Partners

Contact

Bioz Stars

Bioz vStars

Buy from Supplier

Structured Review

Santa Cruz Biotechnology fgf19 shrna
Fgf19 Shrna, supplied by Santa Cruz Biotechnology, used in various techniques. Bioz Stars score: 92/100, based on 2 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
https://www.bioz.com/product/fgf19+shrna/FGF-19+shRNA+(h)+Lentiviral+Particles/pmc05668513-65-3-9
Average 92 stars, based on 2 article reviews

fgf19 shrna - by Bioz Stars, 2026-09

92/100 stars

Images

Plasmid Preparation:

Article Title: FGF 19 genetic amplification as a potential therapeutic target in lung squamous cell carcinomas
Article Snippet: .. The plasmid of FGF19 shRNA (sc‐39480‐V) was purchased from Santa Cruz Biotechnology (Dallas, TX, USA). ..

shRNA:

Image Search Results

Upregulated FGF19 is associated with enhanced self-renewal in HCC. (A - C ) FGF19 expressions in CSCs of Huh-7, RFP/PLC/5 and MHCC97H and corresponding NCSCs: ( A ) mRNA levels were measured by RT-qPCR, ( B ) protein levels were determined by WB, and ( C ) protein concentrations in cell supernatant were measured by ELISA. ( D ) Representative micrographs of FGF19 IHC analysis (400×) in xenograft tumors derived from LCSCs and NCSCs of Huh-7, RFP/PLC/5 and MHCC97H. ( E ) Dynamic expressions of FGF19, Nanog, and ALB were detected in LCSCs at different time points during differentiation. ( F ) Representative micrographs of FGF19, Nanog, Oct-4, and ALB IHC analysis (400×) in 20 HCC samples (left panel); and statistical analysis of integrated optical density (IOD) of FGF19, Nanog, Oct-4, and ALB against immunoglobulin G (right panel). Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: Upregulated FGF19 is associated with enhanced self-renewal in HCC. (A - C ) FGF19 expressions in CSCs of Huh-7, RFP/PLC/5 and MHCC97H and corresponding NCSCs: ( A ) mRNA levels were measured by RT-qPCR, ( B ) protein levels were determined by WB, and ( C ) protein concentrations in cell supernatant were measured by ELISA. ( D ) Representative micrographs of FGF19 IHC analysis (400×) in xenograft tumors derived from LCSCs and NCSCs of Huh-7, RFP/PLC/5 and MHCC97H. ( E ) Dynamic expressions of FGF19, Nanog, and ALB were detected in LCSCs at different time points during differentiation. ( F ) Representative micrographs of FGF19, Nanog, Oct-4, and ALB IHC analysis (400×) in 20 HCC samples (left panel); and statistical analysis of integrated optical density (IOD) of FGF19, Nanog, Oct-4, and ALB against immunoglobulin G (right panel). Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Quantitative RT-PCR, Enzyme-linked Immunosorbent Assay, Derivative Assay

Knockdown of FGF19 attenuates self-renewal in LCSCs. After introducing negative control shRNA (shNC) or shRNA targeting FGF19 (shFGF19) into Huh-7 and RFP/PLC/5 cells, ( A ) RT-qPCR and ( B ) WB were applied to measure levels of FGF19, Nanog, Oct-4, Sox2, ALB and G6P in NCSC transfected with shNC, and in CSCs transfected with shRNA or shFGF19. ( C-F ) The effects of FGF19 silencing on self-renewal features of LCSCs were assessed by ( C ) sphere formation assay, ( D ) clonogenicity assay, ( E ) sorafenib resistance assay, and ( F ) tumorigenic potential assays in vivo . Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: Knockdown of FGF19 attenuates self-renewal in LCSCs. After introducing negative control shRNA (shNC) or shRNA targeting FGF19 (shFGF19) into Huh-7 and RFP/PLC/5 cells, ( A ) RT-qPCR and ( B ) WB were applied to measure levels of FGF19, Nanog, Oct-4, Sox2, ALB and G6P in NCSC transfected with shNC, and in CSCs transfected with shRNA or shFGF19. ( C-F ) The effects of FGF19 silencing on self-renewal features of LCSCs were assessed by ( C ) sphere formation assay, ( D ) clonogenicity assay, ( E ) sorafenib resistance assay, and ( F ) tumorigenic potential assays in vivo . Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Knockdown, Negative Control, shRNA, Quantitative RT-PCR, Transfection, Tube Formation Assay, In Vivo

FGF19 promotes self-renewal via FGFR4 in none-LCSCs. ( A ) RT-PCR and ( B ) WB were applied to measure levels of Nanog, Oct-4, Sox2, ALB and G6P in FGFR4 +/+ NCSCs transfected with empty vector (EV) and FGF19-overexpressing vector (FGF19 OE), as well as in FGFR4 -/- NCSCs. ( C-F ) The effects of exogenous FGF19 expression on self-renewal features of FGFR4 -/- - and FGFR4 +/+ in NCSCs of Huh-7 and RFP/PLC/5 were measured by ( C ) sphere formation assay, ( D ) clonogenicity assay, ( E ) sorafenib resistance assay and, and ( F ) tumorigenic potential assays in vivo . Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: FGF19 promotes self-renewal via FGFR4 in none-LCSCs. ( A ) RT-PCR and ( B ) WB were applied to measure levels of Nanog, Oct-4, Sox2, ALB and G6P in FGFR4 +/+ NCSCs transfected with empty vector (EV) and FGF19-overexpressing vector (FGF19 OE), as well as in FGFR4 -/- NCSCs. ( C-F ) The effects of exogenous FGF19 expression on self-renewal features of FGFR4 -/- - and FGFR4 +/+ in NCSCs of Huh-7 and RFP/PLC/5 were measured by ( C ) sphere formation assay, ( D ) clonogenicity assay, ( E ) sorafenib resistance assay and, and ( F ) tumorigenic potential assays in vivo . Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Transfection, Plasmid Preparation, Expressing, Tube Formation Assay, In Vivo

Enhanced SOCE is critical for FGF19-promoted self-renewal of HCC cells. ( A ) Huh-7 and RFP/PLC/5 NCSCs were pre-treated with DMSO or SKF96365 (5 µM), then Ca 2+ mobilization in Fura-2-loaded cells respectively upon FGF19 (100 ng/ml) stimulation were measured and expressed as means ± SEM of 10 independent cells each group. ( B-D ) The effects of SKF-96365 (5 µM) on self-renewal features of FGF19 (100 ng/ml) treated-NCSCs were measured by ( B ) sphere formation assay, ( C ) clonogenicity assay, ( D ) sorafenib resistance assay. ( E ) RT-PCR and ( F ) WB were applied to measure levels of Nanog, Oct-4, Sox2 and ALB in FGF19 (100 ng/ml) treated NCSCs, in the presence of SKF-96365 (5 µM) or not. ( G ) SOCE related protein levels (STIM1, STIM2, and Orai1); and ( H ) protein levels of p-PLCγ, PLCγ, p-ERK1/2, and ERK1/2 were examined by WB in FGF19 (100 ng/ml) treated- or BSA (100 ng/ml) treated Huh-7 and RFP/PLC/5 NCSCs for 4h. ( I ) Cells were pre-treated with DMSO, 3-NC (20 µM), LY3214996 (2 µM), or 3-NC (20 µM) + LY3214996 (2 µM) for 2h, respectively; then Ca 2+ mobilization in Fura-2-loaded cells respectively upon FGF19 stimulation were measured and expressed as means ± SEM of 12 independent cells each group. Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: Enhanced SOCE is critical for FGF19-promoted self-renewal of HCC cells. ( A ) Huh-7 and RFP/PLC/5 NCSCs were pre-treated with DMSO or SKF96365 (5 µM), then Ca 2+ mobilization in Fura-2-loaded cells respectively upon FGF19 (100 ng/ml) stimulation were measured and expressed as means ± SEM of 10 independent cells each group. ( B-D ) The effects of SKF-96365 (5 µM) on self-renewal features of FGF19 (100 ng/ml) treated-NCSCs were measured by ( B ) sphere formation assay, ( C ) clonogenicity assay, ( D ) sorafenib resistance assay. ( E ) RT-PCR and ( F ) WB were applied to measure levels of Nanog, Oct-4, Sox2 and ALB in FGF19 (100 ng/ml) treated NCSCs, in the presence of SKF-96365 (5 µM) or not. ( G ) SOCE related protein levels (STIM1, STIM2, and Orai1); and ( H ) protein levels of p-PLCγ, PLCγ, p-ERK1/2, and ERK1/2 were examined by WB in FGF19 (100 ng/ml) treated- or BSA (100 ng/ml) treated Huh-7 and RFP/PLC/5 NCSCs for 4h. ( I ) Cells were pre-treated with DMSO, 3-NC (20 µM), LY3214996 (2 µM), or 3-NC (20 µM) + LY3214996 (2 µM) for 2h, respectively; then Ca 2+ mobilization in Fura-2-loaded cells respectively upon FGF19 stimulation were measured and expressed as means ± SEM of 12 independent cells each group. Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Tube Formation Assay, Reverse Transcription Polymerase Chain Reaction

FGF19 signaling facilitates dephosphorylation and nuclear translocation of NFATc2 in non-LCSCs. ( A ) NFAT-RE luciferase activity was measured in FGF19 (100 ng/ml) treated Huh-7 and RFP/PLC/5 NCSCs transfected with si-NC, si-NFATc1, 2, 3, 4, respectively. ( B ) GSEA was performed to evaluate the role of NFATc2 in HCC using available data from TCGA. ( C ) Kaplan-Meier analysis of correlation between the NFATc2 expression and overall survival of HCC patients from TCGA (n=365). ( D and E ) Subcellular location of NFATc2 after FGF19 treatment: Huh-7 NCSCs were pre-treated with DMSO, BLU9931 (100 nM), 3-NC (20 µM), LY3214996 (2 µM), 3-NC (20 µM) + LY3214996 (2 µM), SKF96365 (5 µM), and FK506 (50 nM) for 2h, respectively; then treated with FGF19 (100 ng/ml) for 4h. ( D ) The levels of NFATc2 in cytosol and nucleus, and the phosphorylation level of NFATc2 were determined by WB, Lamin B served as a control of nuclear protein, and α-Tubulin was used as a control of cytoplasmic protein. ( E ) Subcellular location of NFATc2 (green) was detected by immunofluorescence (IF), and cell nucleus was labeled with DAPI (blue). Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: FGF19 signaling facilitates dephosphorylation and nuclear translocation of NFATc2 in non-LCSCs. ( A ) NFAT-RE luciferase activity was measured in FGF19 (100 ng/ml) treated Huh-7 and RFP/PLC/5 NCSCs transfected with si-NC, si-NFATc1, 2, 3, 4, respectively. ( B ) GSEA was performed to evaluate the role of NFATc2 in HCC using available data from TCGA. ( C ) Kaplan-Meier analysis of correlation between the NFATc2 expression and overall survival of HCC patients from TCGA (n=365). ( D and E ) Subcellular location of NFATc2 after FGF19 treatment: Huh-7 NCSCs were pre-treated with DMSO, BLU9931 (100 nM), 3-NC (20 µM), LY3214996 (2 µM), 3-NC (20 µM) + LY3214996 (2 µM), SKF96365 (5 µM), and FK506 (50 nM) for 2h, respectively; then treated with FGF19 (100 ng/ml) for 4h. ( D ) The levels of NFATc2 in cytosol and nucleus, and the phosphorylation level of NFATc2 were determined by WB, Lamin B served as a control of nuclear protein, and α-Tubulin was used as a control of cytoplasmic protein. ( E ) Subcellular location of NFATc2 (green) was detected by immunofluorescence (IF), and cell nucleus was labeled with DAPI (blue). Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: De-Phosphorylation Assay, Translocation Assay, Luciferase, Activity Assay, Transfection, Expressing, Phospho-proteomics, Control, Immunofluorescence, Labeling

Silencing NFATc2 attenuates FGF19-promoted self-renewal in non-LCSCs. ( A-E ) The effects of silencing NFATc2 on FGF19-triggerd self-renewal in Huh-7 NCSCs were evaluated by ( A ) RT-qPCR and ( B ) WB measuring the expressions of Nanog, Oct-4, Sox2, ALB, and G6P, ( C ) sphere formation assay, ( D ) clonogenicity assay and ( E ) sorafenib resistance assay Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: Silencing NFATc2 attenuates FGF19-promoted self-renewal in non-LCSCs. ( A-E ) The effects of silencing NFATc2 on FGF19-triggerd self-renewal in Huh-7 NCSCs were evaluated by ( A ) RT-qPCR and ( B ) WB measuring the expressions of Nanog, Oct-4, Sox2, ALB, and G6P, ( C ) sphere formation assay, ( D ) clonogenicity assay and ( E ) sorafenib resistance assay Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Quantitative RT-PCR, Tube Formation Assay

NFATc2 transcriptionally increase FGF19 expression via binding FGF19 promoter. ( A ) The levels of NFATc2 in cytosol and nucleus from NCSCs and CSCs of HCC cells were determined by WB. NFATc2 and FGF19 mRNA ( B ) and protein ( C ) levels in NCSCs and CSCs of Huh-7 cells were transfected with siNC or si-NFATc2. RT-qPCR ( D ) and WB ( E ) to assess NFATc2 and FGF19 expressions in mock- and NFATc2 OE-Huh-7 NCSCs. ( F ) Representative micrographs (400×) of IHC analysis NFATc2 and FGF19 in 20 HCC samples, and analysis correlation between the IOD of NFATc2 and FGF19 against IgG. ( G ) Bioinformatics analysis predicted four NFAT-REs in the promoter of human FGF19 . ( H ) Luciferase activity assay of FGF19 promoter in Mock- and NFATc2 OE- NCSCs were determined. ( I ) Luciferase activities of wildtype FGF19 promoter (WT) and FGF19 promoter containing single mutant NFAT-RE (M1 to 4) in Mock- or NFATc2 OE- Huh-7 NCSCs were measured. ( J ) ChIP assay of NFATc2 and FGF19 promoter, representative agarose gel results showing recruitment of NFATc2 to the region containing the 4 th NFAT-RE, and IgG used as a negative control. ( K ) CpG island prediction results showed that there are 3 CpG islands in human FGF19 promoter. Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: NFATc2 transcriptionally increase FGF19 expression via binding FGF19 promoter. ( A ) The levels of NFATc2 in cytosol and nucleus from NCSCs and CSCs of HCC cells were determined by WB. NFATc2 and FGF19 mRNA ( B ) and protein ( C ) levels in NCSCs and CSCs of Huh-7 cells were transfected with siNC or si-NFATc2. RT-qPCR ( D ) and WB ( E ) to assess NFATc2 and FGF19 expressions in mock- and NFATc2 OE-Huh-7 NCSCs. ( F ) Representative micrographs (400×) of IHC analysis NFATc2 and FGF19 in 20 HCC samples, and analysis correlation between the IOD of NFATc2 and FGF19 against IgG. ( G ) Bioinformatics analysis predicted four NFAT-REs in the promoter of human FGF19 . ( H ) Luciferase activity assay of FGF19 promoter in Mock- and NFATc2 OE- NCSCs were determined. ( I ) Luciferase activities of wildtype FGF19 promoter (WT) and FGF19 promoter containing single mutant NFAT-RE (M1 to 4) in Mock- or NFATc2 OE- Huh-7 NCSCs were measured. ( J ) ChIP assay of NFATc2 and FGF19 promoter, representative agarose gel results showing recruitment of NFATc2 to the region containing the 4 th NFAT-RE, and IgG used as a negative control. ( K ) CpG island prediction results showed that there are 3 CpG islands in human FGF19 promoter. Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001, NS represents no significant difference.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Expressing, Binding Assay, Transfection, Quantitative RT-PCR, Luciferase, Activity Assay, Mutagenesis, Agarose Gel Electrophoresis, Negative Control

Dual-knockdown of FGF19 and NFATc2 potently represses self-renewal of LCSCs. ( A ) Kaplan-Meier analysis of correlation between the expression of FGF19 and NFATc2 with overall survival of HCC patients from TCGA (n = 365). ( B-F ) Huh-7 and RFP/PLC/5 CSCs were transfected with shNC, either shFGF19, or shNFATc2, shRNAs containing both shFGF19 and shNFATc2 (D-shFGF19 + NFATc2). ( B ) RT-PCR and ( C ) WB were applied to measure levels of FGF19, NFATc2, Nanog, Oct-4, and ALB. The effects of dual-silencing of FGF19 and NFATc2 on self-renewal features of LCSCs in Huh-7 and RFP/PLC/5 were assessed by ( D ) sphere formation assay, ( E ) clonogenicity assay, and ( F ) tumorigenic potential assays in vivo . ( G ) Kaplan-Meier survival curve of orthotopic liver tumor by intrahepatic implantation of Huh-7 LCSCs infected with transfected with shNC, either shFGF19, or shNFATc2, D-shFGF19 + NFATc2 (n = 8/group). Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Journal: Theranostics

Article Title: FGF19/SOCE/NFATc2 signaling circuit facilitates the self-renewal of liver cancer stem cells

doi: 10.7150/thno.56369

Figure Lengend Snippet: Dual-knockdown of FGF19 and NFATc2 potently represses self-renewal of LCSCs. ( A ) Kaplan-Meier analysis of correlation between the expression of FGF19 and NFATc2 with overall survival of HCC patients from TCGA (n = 365). ( B-F ) Huh-7 and RFP/PLC/5 CSCs were transfected with shNC, either shFGF19, or shNFATc2, shRNAs containing both shFGF19 and shNFATc2 (D-shFGF19 + NFATc2). ( B ) RT-PCR and ( C ) WB were applied to measure levels of FGF19, NFATc2, Nanog, Oct-4, and ALB. The effects of dual-silencing of FGF19 and NFATc2 on self-renewal features of LCSCs in Huh-7 and RFP/PLC/5 were assessed by ( D ) sphere formation assay, ( E ) clonogenicity assay, and ( F ) tumorigenic potential assays in vivo . ( G ) Kaplan-Meier survival curve of orthotopic liver tumor by intrahepatic implantation of Huh-7 LCSCs infected with transfected with shNC, either shFGF19, or shNFATc2, D-shFGF19 + NFATc2 (n = 8/group). Data are expressed as means ± SEM (n = 3). *p < 0.05, **p < 0.01, ***p < 0.001.

Article Snippet: The recombinant lentiviral plasmids containing human FGF19 or NFATc2, and shRNA-targeting FGF19 or NFATc2 were purchased from GeneCopoeia (Rockville, MD, USA).

Techniques: Knockdown, Expressing, Transfection, Reverse Transcription Polymerase Chain Reaction, Tube Formation Assay, In Vivo, Infection

Co-amplification and co-overexpression of FGF19 and CCND1 in human LUSC. (A) Copy number amplification distribution of FGF19 and CCND1 in our LUSC cohort # (n = 37). Co-amplification of CCND1 is common in FGF19-amplified LUSC patient in our LUSC cohort (5/9, 55.6%). #Our previous work had summarized all the clinical pathological characteristics of the samples . (B) Amplification of FGF19 and CCND1 is common in TCGA provisional LUSC cohort. Data were shown as mean ± SD bars and compared by unpaired t -test. ** p < 0.01 and **** p < 0.0001. (C) Co-amplification of CCND1 is common in LUSC patients in the TCGA provisional LUSC cohort, that is, 12.7% in Nature 2012, 14.0% Firehose Legacy, 12.4% PanCancer Atlas. (D) FGF19 and CCND1 gene amplification were significantly correlated with their respective expressions. (E) Linear regression and Pearson correlation of mRNA levels between FGF19 and CCND1 in the TCGA provisional LUSC cohort. Positive correlation between expression of FGF19 and CCND1 was observed. (F) Statistical analysis on the individual level based on the TCGA database ( http://www.cbioportal.org ). (a) Amplification and mRNA expression distribution of FGF19 and CCND1. (b, c) A 2 × 4 contingency table was generated for 2 categorical variables, FGF19 and CCND1, in 2 observation states, mRNA high and mRNA no alteration (NA); (b) Each entry represents the number of observations for a given gene in 1 observation state; (c) Entry number of observations of FGF19 and CCND1 in 1 observation state under the premise of co-amplification. (G) Positive correlation between FGF19 and CCND1 protein expression levels in the 16 LUSC tissue samples. Pearson χ 2 test, P <0.001. Scale bar: 100 µm.

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: Co-amplification and co-overexpression of FGF19 and CCND1 in human LUSC. (A) Copy number amplification distribution of FGF19 and CCND1 in our LUSC cohort # (n = 37). Co-amplification of CCND1 is common in FGF19-amplified LUSC patient in our LUSC cohort (5/9, 55.6%). #Our previous work had summarized all the clinical pathological characteristics of the samples . (B) Amplification of FGF19 and CCND1 is common in TCGA provisional LUSC cohort. Data were shown as mean ± SD bars and compared by unpaired t -test. ** p < 0.01 and **** p < 0.0001. (C) Co-amplification of CCND1 is common in LUSC patients in the TCGA provisional LUSC cohort, that is, 12.7% in Nature 2012, 14.0% Firehose Legacy, 12.4% PanCancer Atlas. (D) FGF19 and CCND1 gene amplification were significantly correlated with their respective expressions. (E) Linear regression and Pearson correlation of mRNA levels between FGF19 and CCND1 in the TCGA provisional LUSC cohort. Positive correlation between expression of FGF19 and CCND1 was observed. (F) Statistical analysis on the individual level based on the TCGA database ( http://www.cbioportal.org ). (a) Amplification and mRNA expression distribution of FGF19 and CCND1. (b, c) A 2 × 4 contingency table was generated for 2 categorical variables, FGF19 and CCND1, in 2 observation states, mRNA high and mRNA no alteration (NA); (b) Each entry represents the number of observations for a given gene in 1 observation state; (c) Entry number of observations of FGF19 and CCND1 in 1 observation state under the premise of co-amplification. (G) Positive correlation between FGF19 and CCND1 protein expression levels in the 16 LUSC tissue samples. Pearson χ 2 test, P <0.001. Scale bar: 100 µm.

Article Snippet: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Amplification, Over Expression, Expressing, Generated

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.

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: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

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

FGF19 enhances CCND1-induced inactivation of RB. (A) rhFGF19 phosphorylated RB and promoted expression of CCND1 in SK-MES-1 and H520 cells in a time-dependent manner. (B) pRB and CCND1levels in FGF19 overexpression (SK-MES-1 LV-FGF19) and control (LV-NC) LUSC cells. (C) SK-MES-1 LV-FGF19 cells was transduced with CCND1-knockdown lentivirus (LV-shCCND1), or control lentivirus (LV-shRNA-NC) to construct stable cell lines and quantification of CCND1 in forms as cellular protein and mRNA. (D) pRB and CCND1 levels in FGF19 overexpression and CCND1-knockdown cells (SK-MES-1 LV-FGF19-shCCND1) and control (SK-MES-1 LV-NC-shRNA-NC) LUSC cells. (E) pRB and CCND1 levels in CCND1 knockdown cells (H520-shCCND1) and control (H520-shRNA-NC) LUSC cells. (F) pRB levels in FGF19-knockdown cells (H520-shFGF19) and control (H520-shRNA-NC) LUSC cells, treated with BLU9931, palbociclib, BLU9931 & palbociclib, or DMSO. Right panel: quantifications of pRB. All the data were shown as the mean ± SD. * P < 0.05; ** P < 0.01; *** p < 0.001; **** p < 0.0001; ns, not significant.

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-induced inactivation of RB. (A) rhFGF19 phosphorylated RB and promoted expression of CCND1 in SK-MES-1 and H520 cells in a time-dependent manner. (B) pRB and CCND1levels in FGF19 overexpression (SK-MES-1 LV-FGF19) and control (LV-NC) LUSC cells. (C) SK-MES-1 LV-FGF19 cells was transduced with CCND1-knockdown lentivirus (LV-shCCND1), or control lentivirus (LV-shRNA-NC) to construct stable cell lines and quantification of CCND1 in forms as cellular protein and mRNA. (D) pRB and CCND1 levels in FGF19 overexpression and CCND1-knockdown cells (SK-MES-1 LV-FGF19-shCCND1) and control (SK-MES-1 LV-NC-shRNA-NC) LUSC cells. (E) pRB and CCND1 levels in CCND1 knockdown cells (H520-shCCND1) and control (H520-shRNA-NC) LUSC cells. (F) pRB levels in FGF19-knockdown cells (H520-shFGF19) and control (H520-shRNA-NC) LUSC cells, treated with BLU9931, palbociclib, BLU9931 & palbociclib, or DMSO. Right panel: quantifications of pRB. All the data were shown as the mean ± SD. * P < 0.05; ** P < 0.01; *** p < 0.001; **** p < 0.0001; ns, not significant.

Article Snippet: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Expressing, Over Expression, Control, Transduction, Knockdown, shRNA, Construct, Stable Transfection

FGF19 combined with CCND1 promotes cell cycle progression. (A) rhFGF19 promoted the cell cycle of LUSC cells from G1 to S phase. (B) Enrichment plots of regulation of transcription involved in G1/S transition of mitotic cell cycle and E2F targets signatures according to FGF19 expression levels in an LUSC cohort. (C) GO analysis of cell cycle-related pathways significantly regulated by high expression of FGF19 in LUSC cohort. (D) Representative histograms depicting cell cycle profiles of SK-MES-1 LV-FGF19 and H520 cells with or without BLU9931 and palbociclib. Right panel: quantifications of the histograms. (E) H520 was treated with DMSO, 1 μM BLU9931, 1 μM palbociclib or their combination. (a) Enrichment plot showing HALLMARK_E2F_TARGETS signatures and (b) Heatmap showing the expression of common E2F target genes according to the drug treatment. All the data were shown as the mean ± SD. *** p < 0.001; **** p < 0.0001.

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 combined with CCND1 promotes cell cycle progression. (A) rhFGF19 promoted the cell cycle of LUSC cells from G1 to S phase. (B) Enrichment plots of regulation of transcription involved in G1/S transition of mitotic cell cycle and E2F targets signatures according to FGF19 expression levels in an LUSC cohort. (C) GO analysis of cell cycle-related pathways significantly regulated by high expression of FGF19 in LUSC cohort. (D) Representative histograms depicting cell cycle profiles of SK-MES-1 LV-FGF19 and H520 cells with or without BLU9931 and palbociclib. Right panel: quantifications of the histograms. (E) H520 was treated with DMSO, 1 μM BLU9931, 1 μM palbociclib or their combination. (a) Enrichment plot showing HALLMARK_E2F_TARGETS signatures and (b) Heatmap showing the expression of common E2F target genes according to the drug treatment. All the data were shown as the mean ± SD. *** p < 0.001; **** p < 0.0001.

Article Snippet: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Expressing

CCND1 is essential for FGF19 signaling-mediated LUSC proliferation. (A) Western blot analysis showing protein levels of FGF19 and PCNA in SK-MES-1, H520 and H1703 cells after lentivirus transfection. (B) Clone formation assay of SK-MES-1 cells with or without FGF19 overexpression. (C) Cell proliferation was measured by CCK8 assay in SK-MES-1 LV-FGF19 cells with or without CCND1 knockdown. (D) Clone formation assay of SK-MES-1 LV-FGF19 cells with or without CCND1 knockdown; cultures were stained with crystal violet. (E–J) Data of orthotopic lung cancer model. Cell suspension of SK-MES-1 LV-NC/LV-FGF19/LV-FGF19-shRNA-NC/LV-FGF19-shCCND1 (2 × 10 6 cells) in a total volume of 50 μL mixed with Matrigel (Matrigel: PBS = 1: 4) were injected into the left lung of 5-week-old male BALB/C nude mice (N = 13 mice per group). (E) Experimental timeline for the animal experiment. (F) Representative bioluminescent images (BLI) of the different groups are shown 25 days after orthotopic implantation. Right panel: quantifications of the total flux. (G) Comparison of orthotopic lung cancer models. Tumor was indicated by the arrow. (H) Representative H&E and PCNA staining images of lung samples from each group. (I) Body weight change and (J) overall survival time of indicated groups of nude mice were showed. (K) Higher FGF19 and lower CCND1 mRNA levels are associated with longer overall survival. Data were showed as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. H&E, hematoxylin and eosin.

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: CCND1 is essential for FGF19 signaling-mediated LUSC proliferation. (A) Western blot analysis showing protein levels of FGF19 and PCNA in SK-MES-1, H520 and H1703 cells after lentivirus transfection. (B) Clone formation assay of SK-MES-1 cells with or without FGF19 overexpression. (C) Cell proliferation was measured by CCK8 assay in SK-MES-1 LV-FGF19 cells with or without CCND1 knockdown. (D) Clone formation assay of SK-MES-1 LV-FGF19 cells with or without CCND1 knockdown; cultures were stained with crystal violet. (E–J) Data of orthotopic lung cancer model. Cell suspension of SK-MES-1 LV-NC/LV-FGF19/LV-FGF19-shRNA-NC/LV-FGF19-shCCND1 (2 × 10 6 cells) in a total volume of 50 μL mixed with Matrigel (Matrigel: PBS = 1: 4) were injected into the left lung of 5-week-old male BALB/C nude mice (N = 13 mice per group). (E) Experimental timeline for the animal experiment. (F) Representative bioluminescent images (BLI) of the different groups are shown 25 days after orthotopic implantation. Right panel: quantifications of the total flux. (G) Comparison of orthotopic lung cancer models. Tumor was indicated by the arrow. (H) Representative H&E and PCNA staining images of lung samples from each group. (I) Body weight change and (J) overall survival time of indicated groups of nude mice were showed. (K) Higher FGF19 and lower CCND1 mRNA levels are associated with longer overall survival. Data were showed as the mean ± SD. * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001. H&E, hematoxylin and eosin.

Article Snippet: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Western Blot, Transfection, Tube Formation Assay, Over Expression, CCK-8 Assay, Knockdown, Staining, Suspension, shRNA, Injection, Comparison

Enhanced effects of combined BLU9931 and palbociclib in LUSC cells in vitro . (A) Cells were plated at a low density in 6-well plates, treated with DMSO, BLU9931, palbociclib, or a combination of the two compounds for 7–10 days. (B) CCK8 cell viability assay, (C) Intracellular lactate dehydrogenase (LDH) assay, (D) flow cytometry analysis and (E) β-galactosidase staining assay of H520 and SK-MES-1 LV-FGF19 cells treated with a single agent (BLU9931 or palbociclib) or a combination of both compounds at a fixed ratio (1:1). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; 100 µm.

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: Enhanced effects of combined BLU9931 and palbociclib in LUSC cells in vitro . (A) Cells were plated at a low density in 6-well plates, treated with DMSO, BLU9931, palbociclib, or a combination of the two compounds for 7–10 days. (B) CCK8 cell viability assay, (C) Intracellular lactate dehydrogenase (LDH) assay, (D) flow cytometry analysis and (E) β-galactosidase staining assay of H520 and SK-MES-1 LV-FGF19 cells treated with a single agent (BLU9931 or palbociclib) or a combination of both compounds at a fixed ratio (1:1). * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; 100 µm.

Article Snippet: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

Techniques: In Vitro, Viability Assay, Lactate Dehydrogenase Assay, Flow Cytometry, Staining

Enhanced effects of combined BLU9931 and palbociclib in the animal model of LUSC cells with endogenously or exogenously high expression of FGF19. SK-MES-1 LV-FGF19 (exogenous FGF19 highly expressed LUSC cells) cells or H520 (endogenous FGF19 highly expressed LUSC cells) cells in a volume of 50 μl were subcutaneously injected into the right flanks of BALB/c nude mice. When the tumor reached a volume of 100–200 mm 3 , mice were randomly grouped and orally treated with vehicle, BLU9931 (30 mg/kg, twice a day), palbociclib (100 mg/kg/d), or both drugs in combination for 3 weeks. Tumor volumes were measured every 3 days after the onset of treatment. (A) The tumors were dissected from the mice. (a) SK-MES-1 LV-FGF19 xenograft tumors and (b) H520 xenograft tumors. Growth curve for (c) SK-MES-1 LV-FGF19 and (d) H520 xenograft tumors. (B) Representative histograms depicting cell cycle profiles of tumor cells and quantifications of the histograms. (a) SK-MES-1 LV-FGF19 xenograft tumors; (b) H520 xenograft tumors. (C) The expression of pRB and PCNA in (a) SK-MES-1 LV-FGF19 xenograft tumors and (b) H520 xenograft tumors were detected by western blot. (D) HE and immunofluorescence staining with pRb Ser807/811 , cleaved caspase 3 (Cl. Cas3) and Ki67 of tumor sections. (a) SK-MES-1 LV-FGF19 xenograft tumors; (b) H520 xenograft tumors. Data were shown as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 as calculated by the two-way ANOVA test. Scale bar: 50 µm.

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: Enhanced effects of combined BLU9931 and palbociclib in the animal model of LUSC cells with endogenously or exogenously high expression of FGF19. SK-MES-1 LV-FGF19 (exogenous FGF19 highly expressed LUSC cells) cells or H520 (endogenous FGF19 highly expressed LUSC cells) cells in a volume of 50 μl were subcutaneously injected into the right flanks of BALB/c nude mice. When the tumor reached a volume of 100–200 mm 3 , mice were randomly grouped and orally treated with vehicle, BLU9931 (30 mg/kg, twice a day), palbociclib (100 mg/kg/d), or both drugs in combination for 3 weeks. Tumor volumes were measured every 3 days after the onset of treatment. (A) The tumors were dissected from the mice. (a) SK-MES-1 LV-FGF19 xenograft tumors and (b) H520 xenograft tumors. Growth curve for (c) SK-MES-1 LV-FGF19 and (d) H520 xenograft tumors. (B) Representative histograms depicting cell cycle profiles of tumor cells and quantifications of the histograms. (a) SK-MES-1 LV-FGF19 xenograft tumors; (b) H520 xenograft tumors. (C) The expression of pRB and PCNA in (a) SK-MES-1 LV-FGF19 xenograft tumors and (b) H520 xenograft tumors were detected by western blot. (D) HE and immunofluorescence staining with pRb Ser807/811 , cleaved caspase 3 (Cl. Cas3) and Ki67 of tumor sections. (a) SK-MES-1 LV-FGF19 xenograft tumors; (b) H520 xenograft tumors. Data were shown as mean ± SD and * p < 0.05, ** p < 0.01, *** p < 0.001 and **** p < 0.0001 as calculated by the two-way ANOVA test. Scale bar: 50 µm.

Article Snippet: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

Techniques: Animal Model, Expressing, Injection, Western Blot, Immunofluorescence, Staining

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.

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: FGF19 shRNA lentiviruses and CCND1 shRNA were constructed by the GENECHEM Biotechnology Co. Ltd. (Shanghai, China).

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

Review

fgf19 shrna (Santa Cruz Biotechnology)

Structured Review

Images

Related Articles

Plasmid Preparation:

shRNA:

Similar Products

Image Search Results