Journal: Experimental cell research

Article Title: Scavenger receptor MARCO contributes to macrophage phagocytosis and clearance of tumor cells.

doi: 10.1016/j.yexcr.2021.112862

Figure Lengend Snippet: Fig. 5. MARCO binds integrin on tumor cells. (A) Adhesion and phagocytosis ability of macrophages to SL4 and CT26 cells were determined by co-culture ex periments. (BCD) Expression of integrin α5, β3 and β5 mRNA in SL4 and CT26 tumor cells was determined by RT-qPCR. (E) The interaction between MARCO and integrin was verified by Co-IP experiments. (F) Tumor cells treated with integrin inhibitors were co-cultured with macrophages, and phagocytosis was observed under an inverted microscope. Scale bar = 200 μm (G) The number of phagocytic cells in each group was analyzed using ImageJ. (H) Normal colonic epithelial cells NCM 460 (red) were co-cultured with macrophages (green) for 2 h, and visualized using a confocal microscope. Scale bar = 50 μm. Calcein-AM, tumor cells SL4 and CT26; Ctrl, SL4 co-culture with macrophage; Cilengitide, SL4 treated with integrin inhibitors Cilengitide co-culture with macrophage; Mito-red, NCM 460; GFP, macrophages. Data are representative of 3 independent experiments; Data are shown as mean ± SEM from triplicate and quadruplicate values. * denotes p < 0.05.

Article Snippet: Tumor cells were precultured with or without the integrin inhibitor Cilengitide, at a 10 μM concentration (TargetMol, Catalog#T2494), for 2 h. Fluorescent labelling of tumor cells was performed as described above, and tumor cells were co-cultured with macrophages for 2 h. Cells that do not adhere were washed away, and the culture was then inspected under an inverted fluorescence microscope.

Techniques: Co-Culture Assay, Expressing, Quantitative RT-PCR, Co-Immunoprecipitation Assay, Cell Culture, Inverted Microscopy, Microscopy

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: Journal of the American Chemical Society

Article Title: A Chemically Cross-Linked Knottin Dimer Binds Integrins with Picomolar Affinity and Inhibits Tumor Cell Migration and Proliferation

doi: 10.1021/ja508416e

Figure Lengend Snippet: Figure 1. Binding assays. Competition binding of Alexa488-2.5F monomer to integrin receptors expressed on U87MG cells (A) and MDA-MB-231 cells (B) by knottin monomer, dimers, and Cilengitide, as measured by

Article Snippet: 2 × 104 cells were incubated with varying concentrations of knottins or Cilengitide (BOC Sciences) and a constant concentration of competitor for 10 h at 4 °C (Supporting Information).

Techniques: Binding Assay

Journal: Journal of the American Chemical Society

Article Title: A Chemically Cross-Linked Knottin Dimer Binds Integrins with Picomolar Affinity and Inhibits Tumor Cell Migration and Proliferation

doi: 10.1021/ja508416e

Figure Lengend Snippet: Figure 2. Effect of D2.5F_1 dimer (red), 2.5F monomer (blue), or Cilengitide (green) on U87MG cell migration compared to untreated control (brown). (A) Bright field images showing tumor cell migration into

Article Snippet: 2 × 104 cells were incubated with varying concentrations of knottins or Cilengitide (BOC Sciences) and a constant concentration of competitor for 10 h at 4 °C (Supporting Information).

Techniques: Migration, Control

Journal: Journal of the American Chemical Society

Article Title: A Chemically Cross-Linked Knottin Dimer Binds Integrins with Picomolar Affinity and Inhibits Tumor Cell Migration and Proliferation

doi: 10.1021/ja508416e

Figure Lengend Snippet: Figure 3. Effect of D2.5F_1 dimer (red), 2.5F monomer (blue), or Cilengitide (green) on MDA-MB-231 cell

Article Snippet: 2 × 104 cells were incubated with varying concentrations of knottins or Cilengitide (BOC Sciences) and a constant concentration of competitor for 10 h at 4 °C (Supporting Information).

Techniques:

Journal: Chinese Journal of Cancer

Article Title: Standards of care and novel approaches in the management of glioblastoma multiforme

doi: 10.5732/cjc.013.10207

Figure Lengend Snippet: MicroRNAs (miRNAs) associated with epithelial-mesenchymal transition (EMT)

Article Snippet: Cilengitide (Merck KGaA, Darmstadt, Germany) is a selective inhibitor of these integrins and has been extensively evaluated in GBM.

Techniques: