Journal: Cancer Research
Article Title: Omental Preadipocytes Stimulate Matrix Remodeling and IGF Signaling to Support Ovarian Cancer Metastasis
doi: 10.1158/0008-5472.CAN-23-2613
Figure Lengend Snippet: IGF1 is elevated in coculture and regulates ECM gene expression and alternative NF-κB. A, Representative adipokine secretory profiles from ACI-23: preadipocyte cocultures and monoculture controls in complete media. B, Quantification of IGFBPs from adipokine arrays. One-way ANOVA with Tukey post hoc test ( n = 4). C, IGF1 ELISA of supernatants from 6-day preadipocyte and cancer cell cocultures, with monoculture controls in complete media. Two-way ANOVA with Sidak post hoc test ( n = 3). D, IGF1 ELISA of supernatants from preadipocyte and cancer cell cocultures, with monoculture controls used for RNA sequencing, all in complete media. Two-way ANOVA with Sidak post hoc test ( n = 3). E, Gene expression of IGF1 (top) and IGF2 (bottom) in primary and omental tumors from three publicly available data sets relativized to the average of the primary tumors. Unpaired t test. F, ECM gene expression via qRT-PCR of ACI-23 cells treated with 100-ng/mL IGF1 for 72 hours in complete media; data are fold change of vehicle. Unpaired t test ( n = 3–5). G, NF-κB activity in OVCAR8-NF-κB-Luc cells treated with vehicle control or 100-ng/mL IGF1 for 4, 24, 48, and 72 hours in complete media. One-way ANOVA with Tukey post hoc ( n = 3). H, Gene expression of RELA and RELB in ACI-23 treated with vehicle control or 100-ng/mL IGF1 for 72 hours in complete media measured via qRT-PCR. Unpaired t test ( n = 3). I, RelB expression in cytosolic and nuclear fractions of ACI-23 cells treated with IGF1 for 72 hours in SFM. Lysate fractions were run on the same gel but displayed as separate panels for clarity. Western blots were also probed for nuclear and cytosolic housekeeping proteins lamin A/C, GAPDH, and α-tubulin ( n = 2). J, NF-κB activity in OVCAR8-NF-κB-Luc cells treated with SFM or CM for 4, 24, 48, and 72 hours. One-way ANOVA with Tukey post hoc ( n = 3). K, Gene expression of RELA and RELB in ACI-23 cocultured with preadipocytes for 72 hours in complete media measured via qRT-PCR. Unpaired t test ( n = 3). *, P < 0.05; **, P < 0.01; ****, P < 0.0001; ns, not significant.
Article Snippet: Secreted factors from cocultures and monoculture controls were measured using the Proteome Profiler Adipokine Array (ARY024) from R&D Systems according to manufacturer’s instructions.
Techniques: Expressing, Enzyme-linked Immunosorbent Assay, RNA Sequencing Assay, Quantitative RT-PCR, Activity Assay, Control, Western Blot
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