Fig 2: Detection of inflammation and oxidative-reduction related factors in heart tissue of mice using ELISA method (A) iNOS, (B) COX-2, (C) CCL9, (D) CXCL1, (E) CXCL9 and (F) CXCL11 in mouse heart tissues. Data are presented as the mean ± SD. Each experiment was repeated three times independently. *P<0.05 vs. NC group. #P<0.05 vs. OT group. DHE, dehydrocostus lactone; TXNIP, thioredoxin-interacting protein; NC, negative control; OT, DHE treatment group; OI, DHE treatment combined with TXNIP inhibition group; OH, DHE treatment combined with TXNIP overexpression group; iNOS, inducible nitric oxide synthase; COX-2, cyclooxygenase-2; CCL9, C-C motif ligand 9; CXCL, chemokine C-X-C motif ligand.

Fig 3: Overview of obesity and radiation associated adipose dysfunction promotes a pro-tumorigenic prostate microenvironment. This stress-adapted phenotype is characterized by enhanced survival signaling, oxidative stress, chronic inflammation, senescence-associated remodeling, and ECM remodeling, collectively establishing a tumor-permissive stromal niche. The obese and irradiated obese adipose microenvironment differentiates prostate fibroblasts toward myofibroblasts and senescent fibroblasts. In turn, myofibroblasts or senescent fibroblasts under obese and irradiated obese adipose microenvironments secrete pro-inflammatory chemokines, including CXCL10 and CXCL11, which promote prostate cancer migration through CXCR3-mediated paracrine signaling. Radiation exposure sustains obesity-associated pathological state within the prostate microenvironment and prostate fibroblasts.

Fig 4: Prostate fibroblasts exposed to an obese or irradiated obese adipose microenvironment promote prostate cancer progression. (A) Migration assay of PC3 and C42B toward the MPF exposed to obese and irradiated obese 3T3-L1 adipocyte CM and normal MPF CM for 48 h. (B) Cell viability of PC3 and C42B when exposed to MPF CM incubated in obese and irradiated obese 3T3-L1 adipocytes for 48 h. (C) Top, Western blot analysis of the cell survival marker, Bcl-2, on C42B cells incubated with normal MPF CM and MPF CM exposed to obese and irradiated obese 3T3-L1 adipocytes for 48 h. Ponceau staining was used as the loading control. Bottom, densitometry of immunoblot showing Bcl-2 expression normalized to the corresponding Ponceau staining. Supplementary Figure S6 represents the original Western blots. (D) Western blot analysis of EMT markers and p-Akt in C42B cells after 48 h of exposure to CM from MPFs incubated in obese and irradiated obese 3T3-L1 adipocytes and from normal MPFs. Ponceau was utilized as loading control. Supplementary Figure S7 represents the original membranes of Western blot. (E) Densitometry of EMT markers and p-Akt in C42B normalized to ponceau. (F) Cytokine array of culture supernatants collected from the MPF exposed to obese and irradiated obese 3T3-L1 adipocytes for 96 h, with media replenishment at 48 h. After 96 h, the CM from obese and irradiated obese adipocytes was removed from MPFs from the respective conditions and washed with PBS 3 times. After 24 h of culture, the fresh MPF medium was collected from the respective treated conditions and normal conditions n = 1. The original membrane of cytokine array is represented in Supplementary Figure S8. (G) CXCL11 and CXCL10 ELISAs performed on the CM collected from normal MPF and fibroblasts exposed to obese and irradiated obese 3T3-L1 adipocytes. (H) Transwell migration assay of PC3 and C42B demonstrating the functional role of CXCR3 signaling in PCa migration toward the MPF exposed to obese and irradiated obese 3T3-L1 adipocytes. All in vitro data are presented as mean ± SEM from four or more biological independent experiments except (F). PCa cells incubated in standard RPMI culture served as a control. Statistical significance was determined by one-way ANOVA followed by a Tukey’s multiple-comparison test. * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001.