Journal: International Journal of Biological Sciences

Article Title: Integrative Single-Cell and Spatial Transcriptomics Analysis Reveals ECM-remodeling Cancer-associated Fibroblast-Derived POSTN as a Key Mediator in Pancreatic Ductal Adenocarcinoma Progression

doi: 10.7150/ijbs.108618

Figure Lengend Snippet: CAF-derived POSTN induces the translation to EMT-subtype via PI3K/AKT/β-catenin signaling in PDAC Cells. (A) Differential gene expression analysis of ductal cells based on fibroblast-derived POSTN levels. Tumor samples from the CRA001160 dataset were classified into high- and low- POSTN groups (75th percentile cutoff), and ductal cell gene expression profiles were compared to identify differences linked to CAF-derived POSTN. (B) GSEA of upregulated genes in ductal cells from POSTN-high samples identified enriched KEGG pathways. (C-F) GSEA of upregulated genes revealed enrichment in the EMT pathway, focal adhesion pathway, PI3K-AKT pathway, and WNT pathway. (G-H) Western blot analysis of β-catenin expression, and phosphorylation of FAK, AKT, and GSK-3β in BxPC-3 and PANC-1 cells after 24-hour treatment with rhPOSTN at varying concentrations. (I) Schematic illustration of how CAF-derived POSTN drives the EMT phenotype in PDAC cells via integrin αvβ5/FAK/PI3K/AKT/β-catenin signaling pathway.

Article Snippet: BxPC-3 cells were pretreated for 24 h with CAF-oePOSTN CM, CAF-NC CM, recombinant human Periostin (rhPOSTN, Novoprotein, CJ39), or integrin αvβ5 receptor inhibitor (MCE, HY-16141).

Techniques: Derivative Assay, Gene Expression, Western Blot, Expressing, Phospho-proteomics

Journal: International Journal of Biological Sciences

Article Title: Integrative Single-Cell and Spatial Transcriptomics Analysis Reveals ECM-remodeling Cancer-associated Fibroblast-Derived POSTN as a Key Mediator in Pancreatic Ductal Adenocarcinoma Progression

doi: 10.7150/ijbs.108618

Figure Lengend Snippet: Integrin αvβ5 inhibitors partially reverse POSTN-induced proliferation, colony formation, migration, and invasion of PDAC cells. (A) Co-localization of POSTN and intergrin β5 in PDAC tissues. Immunofluorescence staining showing POSTN (green), integrin β5 (red), and nuclei (blue) in PDAC patient resection specimens. Scale bars, 25 μm. (B-C) Effect of integrin αvβ5 inhibition on POSTN-induced proliferation in BxPC-3 and PANC-1 cells. Cells were treated with: (1) negative control, (2) 500 ng/mL rhPOSTN, (3) integrin αvβ5 inhibitor (HY-16141) at 1/5 IC 50 concentration, or (4) integrin αvβ5 inhibitor pretreated for 24 hours, followed by 500 ng/mL rhPOSTN. Cell proliferation was assessed using CCK-8 assays. (D) Colony formation assays in BxPC-3 and PANC-1 cells following the same treatments as in (B-C). (E-F) Wound healing assays in BxPC-3 and PANC-1 cells after treatments as described in (B-C), assessing migration capacity. (G-H) Transwell assays in BxPC-3 and PANC-1 cells to assess migation (G) and invasion (H) under the same treatment as in (B-C). (I) Western blot analysis of β-catenin expression, and phosphorylation of FAK, AKT, and GSK-3β in BxPC-3 and PANC-1 cells after 24-hour treatment as described in (B-C).

Article Snippet: BxPC-3 cells were pretreated for 24 h with CAF-oePOSTN CM, CAF-NC CM, recombinant human Periostin (rhPOSTN, Novoprotein, CJ39), or integrin αvβ5 receptor inhibitor (MCE, HY-16141).

Techniques: Migration, Immunofluorescence, Staining, Inhibition, Negative Control, Concentration Assay, CCK-8 Assay, Western Blot, Expressing, Phospho-proteomics

Journal: Oncology Research

Article Title: Ursolic Acid Attenuates TGF-β1-Induced Epithelial–Mesenchymal Transition in NSCLC by Targeting Integrin αVβ5/MMPs Signaling

doi: 10.3727/096504017X15051723858706

Figure Lengend Snippet: Ursolic acid inhibits TGF-β1-induced integrin αVβ5 signaling. (A) The effect of ursolic acid on TGF-β1-induced expression of integrin αVβ5 was evaluated using Western blot analysis. Briefly, H1975 cells were treated with TGF-β1 alone or in combination with ursolic acid for 24 h. Western blotting analysis of integrin αVβ5 expression by H1975 cells. (B) Immunofluorescence revealed that H1975 cells exposed to TGF-β1 significantly increased their expression of integrin αVβ5, while ursolic acid inhibited integrin αVβ5 levels induced by TGF-β1.

Article Snippet: Ursolic acid was dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA) prior to incubation with or without TGF-β1 (Roche, Mannheim, Germany) at a concentration of 5 ng/ml for 24 h. To investigate the effects of TGF-β1 or ursolic acid on cell morphology, an integrin αVβ5 inhibitor SB273005 (Selleck, Houston, TX, USA) was used at 100 nM with or without TGF-β1.

Techniques: Expressing, Western Blot, Immunofluorescence

Journal: Oncology Research

Article Title: Ursolic Acid Attenuates TGF-β1-Induced Epithelial–Mesenchymal Transition in NSCLC by Targeting Integrin αVβ5/MMPs Signaling

doi: 10.3727/096504017X15051723858706

Figure Lengend Snippet: Integrin αVβ5 inhibitor SB273005 blocked the effects of ursolic acid on TGF-β1-induced EMT in H1975 cells. (A) Effects of SB273005 on the migration of H1975 cells with TGF-β1. A cell migration assay was performed by scratching the cell layer prior to drug treatment, and time-lapse images were obtained from 0 to 24 h. H1975 cells were treated with SB273005 (100 nM) alone or ursolic acid combined with SB273005 for 24 h in the presence of TGF-β1. (B) H1975 cells were subjected to Transwell invasion assay in the presence of SB273005 (100 nM) alone or ursolic acid combined with SB273005. Data are presented as the mean ± SD from three independent measurements. ** p < 0.01 versus control.

Article Snippet: Ursolic acid was dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, St. Louis, MO, USA) prior to incubation with or without TGF-β1 (Roche, Mannheim, Germany) at a concentration of 5 ng/ml for 24 h. To investigate the effects of TGF-β1 or ursolic acid on cell morphology, an integrin αVβ5 inhibitor SB273005 (Selleck, Houston, TX, USA) was used at 100 nM with or without TGF-β1.

Techniques: Migration, Cell Migration Assay, Transwell Invasion Assay, Control