Journal: Frontiers in Oncology

Article Title: Growth hormone receptor antagonism downregulates ATP-binding cassette transporters contributing to improved drug efficacy against melanoma and hepatocarcinoma in vivo

doi: 10.3389/fonc.2022.936145

Figure Lengend Snippet: GHRA suppresses syngeneic mouse melanoma growth in vivo . (A) Mouse Fluc-B16-F10 cells grafted intradermally on the right flank of syngeneic C57BL6/J wild-type (WT) and GHA male mice (transgenic for bGH G119K GHR antagonist) (n=6). Tumor growth over 4-weeks was followed by luminescent imaging following luciferin injections. Luciferin signal was quantified and plotted on the right. The changes in tumor volume (Fluc-B16-F10 in WT and GHA mice) from digital caliper measurement (B) and tumor mass (C) corroborate the suppressed tumor growth in GHA mice which has markedly lower serum IGF1 levels (D) due to presence of a circulating GHRA. Western-blot analysis of the GH downstream signaling mediators – phosphorylated STAT5, AKT and SRC kinase in the tumors of GHA and WT mice (E) . B16-F10 cells in culture when treated for 72-hours with serum collected from WT and GHA mice showed suppressed growth rate in the GHA mouse serum (F) . (*p < 0.05, mouse studies – repeated measure using SPSS; cell viability - Students t test, n = 3).

Article Snippet: The sera were then collected and measured by ELISA for IGF1 (#22-IG1MS-E01, ALPCO, Salem, NH, USA) according to the manufacture’s guidelines.

Techniques: In Vivo, Transgenic Assay, Imaging, Western Blot

Journal: Frontiers in Oncology

Article Title: Growth hormone receptor antagonism downregulates ATP-binding cassette transporters contributing to improved drug efficacy against melanoma and hepatocarcinoma in vivo

doi: 10.3389/fonc.2022.936145

Figure Lengend Snippet: Effect of GHRA on response of syngeneic mouse hepatocellular carcinoma tumors to sorafenib treatment in vivo. (A) Mouse Hepa1-6 hepatocellular carcinoma (HCC) cells respond to bovine GH stimulation in culture showing increased cell viability and STAT5 activation after 72 hours (A) as well as increase in the EC50 dose of sorafenib (B) . Additionally, GH treatment increased ABC transporter levels in the cultured Hepa1-6 cells (C) . Mouse Hepa1-6 cells grafted subcutaneously on the right flank of syngeneic C57BL6/J wild-type (WT) and GHA mice (transgenic for bGH G119K GHR antagonist) (n=6). The changes in tumor volume (Hepa1-6 in WT and GHA mice) from digital caliper measurement (D) and post-dissection tumor mass (E) show suppressed tumor growth and improved tumoral response to sorafenib in the GHA mice. (F) Spearman correlation analysis for transcript levels of GHR and ABC transporter and IGF1R and ABC transporters in the tumor of 371 HCC patients in the TCGA cohort. (G) Serum IGF1 levels of Hepa1-6 tumor-bearing GHA mice are significantly lower than that of tumor-bearing WT mice. (H–J) Correlation of overall survival probability of 371 HCC patients in the TCGA cohort with RNA expression of IGF1R (H) , or ABCB1 (I) , or ABCC1 (J) . (*p < 0.05, ***p < 0.001, mouse studies – repeated measure using SPSS; other assays - Students t test, n = 3).

Article Snippet: The sera were then collected and measured by ELISA for IGF1 (#22-IG1MS-E01, ALPCO, Salem, NH, USA) according to the manufacture’s guidelines.

Techniques: In Vivo, Activation Assay, Cell Culture, Transgenic Assay, Dissection, RNA Expression

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) Differential gene expression analysis was performed comparing H2B–EGFP –positive and H2B–EGFP –negative cells within the MC-progenitor cluster identified by single-cell RNA sequencing. Genes are ranked by statistical significance. Foxm1 is significantly enriched in the H2B–EGFP –positive population, whereas Tgfb1 is enriched in the H2B–EGFP –negative population, indicating divergent transcriptional programs associated with Wnt activity. (B) Feature plots were generated to visualize expression of representative genes across mandibular condylar cartilage populations. Wnt-responsive cells show enriched expression of Foxm1 and IGF signaling–related genes ( Igf1 , Igf1r , Igfbp4 , Igfbp7 ), whereas Wnt-low populations express Tgfb1 and related factors ( Igf2r , Igfbp5 , Igfbp6 ), supporting distinct signaling states. (C) Western blot analysis was performed in isolated Wnt-responsive cells transfected with control vector or constitutively active β-catenin (S33Y). Cells were stimulated with recombinant IGF1 for the indicated time points. β-catenin activation enhances Foxm1 expression and downstream mitogenic signaling, including ERK and IGF1R phosphorylation, indicating that β-catenin promotes proliferative signaling responses. (D) Co-immunoprecipitation was performed to assess interaction between β-catenin and Foxm1. Cell lysates immunoprecipitated with anti–β-catenin antibody show enrichment of Foxm1 compared with control IgG, indicating a physical association between β-catenin and Foxm1. (E,F) Histological and immunofluorescence analyses were performed on mandibular condyles from control and Axin2 CreERT2 ;Ctnnb1 fl/+ ;Foxm1 fl/+ compound heterozygous mice at P42. H&E staining reveals reduced fibrocartilage thickness, and Ki67 staining shows decreased proliferative activity, indicating cooperative effects of β-catenin and Foxm1 in maintaining fibrocartilage proliferation. Scale bar, 100 μm. (G) Quantification of fibrocartilage thickness and Ki67-positive cells was performed. Compound heterozygous mice show reduced fibrocartilage thickness and decreased proliferation compared with controls. Data are presented as mean ± s.d. Each dot represents one biologically independent animal. (H,I) Histological and immunofluorescence analyses were performed on mandibular condyles from control and Axin2 CreERT2 ;Foxm1 fl/fl mice at P42. Foxm1 deletion results in marked condylar hypoplasia and reduced proliferative activity, indicating a critical role for Foxm1 in fibrocartilage growth. Scale bar, 100 μm. (J) Quantification of cartilage thickness and proliferative indices was performed in Foxm1 conditional knockout mice. Foxm1 deficiency significantly reduces cartilage growth and proliferation. Data are presented as mean ± s.d. Each dot represents one biologically independent animal. Statistical significance was assessed using two-tailed Student’s t-test. n.s., not significant; **P < 0.01; ****P < 0.0001. Abbreviations: sz, superficial zone; fc, fibrocartilage zone; cc, chondrocartilage zone.

Article Snippet: Cells were stimulated with recombinant mouse IGF1 (Cat No. 791-MG-050, R&D systems) for 0, 30, 60, or 180 min.

Techniques: Gene Expression, Single Cell, RNA Sequencing, Activity Assay, Generated, Expressing, Western Blot, Isolation, Transfection, Control, Plasmid Preparation, Recombinant, Activation Assay, Phospho-proteomics, Immunoprecipitation, Immunofluorescence, Staining, Knock-Out, Two Tailed Test

Journal: bioRxiv

Article Title: A Wnt-responsive fibrocartilage progenitor system coordinates postnatal mandibular condylar cartilage growth

doi: 10.64898/2026.03.25.714159

Figure Lengend Snippet: (A) RNAscope in situ hybridization showing Foxm1 transcript localization within the fibrocartilage compartment of the mandibular condyle. (B) RNAscope detection of Igf1 transcripts enriched in the superficial region of the fibrocartilage layer. (C) Violin plot comparing Foxm1 expression between H2B-EGFP –positive and H2B-EGFP –negative cells within the MC-progenitor cluster. Scale bars: 100 μm.

Article Snippet: Cells were stimulated with recombinant mouse IGF1 (Cat No. 791-MG-050, R&D systems) for 0, 30, 60, or 180 min.

Techniques: RNAscope, In Situ Hybridization, Expressing