Fig. 3: NPs inhibited CAFs-mediated CRC growth and metastasis in vitro by regulating multiple signaling pathways and secreted chemokines. | Nature Communications

Fig. 3: NPs inhibited CAFs-mediated CRC growth and metastasis in vitro by regulating multiple signaling pathways and secreted chemokines.

From: Amelioration of colorectal cancer-associated fibroblasts in immunosuppressive microenvironment by ferroptosis-based nanotherapy

Fig. 3: NPs inhibited CAFs-mediated CRC growth and metastasis in vitro by regulating multiple signaling pathways and secreted chemokines.

a Schematic illustration of cell viability and transwell migration assays using HCT116 cells treated with supernatants from NFs, CAFs, or NPs-treated CAFs. b Transwell migration images presenting the number of migrated HCT116 cells treated with CM from NFs, CAFs, or NPs-treated CAFs for 24 h. c Cell viability of HCT116 cells treated as in (b) by the CCK8 assay. d Western blot of E-cadherin, N-cadherin, and Vimentin expression in HCT116 cells treated with CM from NFs, CAFs, or NPs-treated CAFs for 24 h; β-actin as a loading control. e, f Bubble plots of (e) GO and (f) KEGG pathway enrichment analyses for differentially expressed genes (DEGs) in NPs-treated CAFs compared to controls. g, h MSigDB Hallmark GSEA analysis showing enrichment of (g) chemokine activity and (h) ferroptosis pathways in NPs-treated CAFs compared to controls. ES: enrichment score; NES: normalized enrichment score. i Heatmap and histogram displaying gene expression levels of DEGs (log₂|FC | ≥ 1) from three chemokine-related GO pathways (GO:0016493, GO:0048020, GO:0008009) in NPs-treated CAFs compared to controls, ranked by differential expression. j QRT-PCR quantification of CCL3, CXCL12, and CCR7 expression in NPs-treated CAFs compared to controls. k, l Anti-human ELISA quantification of (k) CCL3 and (l) CXCL12 concentrations in supernatants from CAFs treated without or with NPs for 24 h. m Western blot of MAPK, p-MAPK (Thr180/Tyr182), NF-κB, p-NF-κB (Ser536), AKT, and p-AKT (Ser473) expression in CAFs treated without or with NPs for 24 h; β-actin as a loading control. n = 3 independent experiments in (b, c) and (j–l). Western blot experiments in (d, m) are repeated independently three times with similar results. Data are presented as means ± SD. Statistical analyses are performed using one-way ANOVA with multiple comparisons for (b, c); and two-tailed unpaired Student’s t-test for (j–l). Source data are provided as a Source Data file.

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