Figure 7 from Vitronectin Enrichment in Prostate Cancer Liver Metastases Promotes Adhesion and Survival
Year: 2026
Type: other
Abstract: VTN is associated with greater liver metastatic burden, and its phenotypic effects may be mitigated by defactinib. A, Schematic of the in vivo experimental plan wherein mice undergo orthotopic 22Rv1 injection to the anterior prostate, tail-vein injections of VTN or PBS every other day, and necropsy to diagnose metastatic spread. B, Tumor volume, tumor mass, and liver mass were measured at necropsy. Results are plotted for two biological replicates. C, Whole livers were excised and examined for metastatic foci. Livers were chunked, formalin-fixed, paraffin-embedded, and hematoxylin and eosin (H&E)–stained before re-examination for micrometastases. Whole tissue with grossly visible metastatic foci highlighted in green and H&E of tissue regions with metastases and corresponding AR IHC are shown. D, Cells were plated and serum-starved overnight with or without 1 μmol/L defactinib. Then, cells were exposed to VTN and DMSO or VTN and defactinib for the specified amounts of time before cells were lysed, and immunoblotting was performed. Red triangles indicate the pERK bands. E, Adhesion assays were performed in serum-free media with the specified compounds and either 5 μg/mL VTN in dH2O or dH2O alone. Bars show three biological replicates, and significance was calculated with two-tailed Welch t tests. Complete, complete media used as a positive control; def., defactinib; ns, not statistically significant. F, Cells were plated in 6-well plates in serum-free media with either 5 μg/mL VTN in dH2O or dH2O alone, in the context of the MEK and PI3K inhibitors trametinib and alpelisib in combination or the FAK inhibitor defactinib, and allowed to adhere for 24 hours. Cells were harvested for PI staining followed by flow cytometry, and data were gated to exclude debris and doublets. Hypodiploid cell fractions were calculated as described in “Materials and Methods,” and statistical significance was determined from individual experiments with a two-tailed Fisher exact test. alp., alpelisib; tram., trametinib. *, P ≤ 0.05; **, P ≤ 0.01; ***, P ≤ 0.001.
Subfield: Pulmonary and Respiratory Medicine
Field: Medicine
Domain: Health Sciences
Open Access status: gold
OpenAlex ID: W7213467775
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