human lung adenocarcinoma a549 cells fap negative  (Procell Inc)

Journal: Signal Transduction and Targeted Therapy

Article Title: Breast cancer-associated fibroblast-derived exosomal miR-1290 promotes triple-negative breast cancer cell proliferation

doi: 10.1038/s41392-026-02922-y

Figure Lengend Snippet: miR-1290 is upregulated in breast cancer and BCAF/MB-231-derived exosomes. a Nanoparticle tracking analysis of exosomes isolated from BCAFs and MB-231 cells showing their size distribution. The mean size and standard deviation (SD) are indicated for each sample. b TEM images of exosomes isolated from BCAFs (upper) and MB-231 cells (lower) showing the characteristic morphology of the exosomes. Scale bar, 100 nm. c Western blot analysis of exosome markers (CD63, CD81, and flotillin-1), CAF markers (α-SMA and FAP) and cellular markers (GM130 and H3) in BCAF cells, MB-231 cells and their derived exosomes. d BCAF-derived exosome uptake by TNBC cells. PKH67-labeled BCAF-derived exosomes (green) were incubated with BT-549 and Hs578T cells for 1 hour. The cell nuclei were stained with Hoechst 33342 (blue). Scale bar, 20 μm. e Heatmap showing miRNA expression profiles in exosomes derived from BCAFs and MB-231 cells. Red indicates high expression, and blue indicates low expression. miR-1290 (highlighted in red) is highly expressed in both exosome types. f Volcano plots showing differentially expressed miRNAs between breast cancer or TNBC samples and normal controls in five independent GEO datasets ( GSE37963 , GSE45666 , GSE45498 , GSE38167 , and GSE61438 ). Red and blue dots represent upregulated and downregulated miRNAs, respectively, selected using fold-change and nominal p value cutoffs (|log2(fold change)| > 0.58 and nominal p < 0.05, two-tailed Welch’s t test). g Venn diagram analysis of upregulated miRNAs across the five GEO datasets revealed four commonly upregulated miRNAs. h Violin plot showing miR-1290 expression levels in serum samples from normal individuals ( N , n = 2686) and breast cancer patients ( P , n = 1280) from the GSE73002 dataset. The expression of miR-1290 was significantly upregulated in the breast cancer patient samples (*** p < 0.001, Mann‒Whitney U test). i Analysis of serum exosomal miR-1290 levels in normal ( N , n = 20) and TNBC patient ( P , n = 39) samples collected at Keimyung University Dongsan Hospital. The results support the potential of miR-1290 as a candidate diagnostic marker in an independent cohort (*** p < 0.001, Mann‒Whitney U test). j Kaplan‒Meier plot of overall survival for TNBC patients based on serum exosomal miR-1290 expression levels from Keimyung University Dongsan Hospital ( p = 0.006, log-rank test)

Article Snippet: Membranes were incubated with the following primary antibodies: CD63 (1:200; ABclonal, A5271), CD81 (1:200; ABclonal, A5270), flotillin-1 (1:1000; ABclonal, A6220), α-SMA (1:1000; ABclonal, A17910), FAP (1:1000; ABclonal, A23789 ), GM130 (1:1000; Abcam, ab52649), H3 (1:1000; Cell Signaling Technology, 4620S), ACTB (1:20,000; ABclonal, AC026), TBC1D4 (1:200; Cell Signaling Technology, 2670S), P70S6K (1:500; Cell Signaling Technology, 9202S), phospho-P70S6K (1:500; ABclonal, AP0564), and Ki67 (1:1000; Abcam, ab16667).

Techniques: Derivative Assay, Exosomes, Analysis, Isolation, Standard Deviation, Morphology, Western Blot, Labeling, Incubation, Staining, Expressing, Two Tailed Test, Blood/Plasma/Serum Samples, Diagnostic Assay, Marker

Journal: Acta Pharmaceutica Sinica. B

Article Title: Targeting FAP α -positive hepatic stellate cells ameliorates the formation of pre-metastatic niche

doi: 10.1016/j.apsb.2026.04.018

Figure Lengend Snippet: FAP α + HSCs upregulate NF- κ B/NLRP3/caspase-1/IL-18 signaling pathway. (A) KEGG analysis of the Top100 upregulated genes in LX-2 FAP cells. (B) GO analysis of paracancerous and healthy HSCs from scRNA sequencing. (C) Western blotting analysis of FAP α , NF κ B-p65, p- NF κ B-p65, NLRP3, pro caspase-1, caspase-1 P 20, IL-18 in LX-2 FAP cells with or without SP-13786 ( n = 3). (D) Immunocytochemistry of NLRP3 in LX-2 FAP cells with or without SP-13786 ( n = 3). Scale bar, 10 μm. (E) ELISA analysis of IL-18 concentration in LX-2 FAP cells ( n = 3). (F) ELISA analysis of IL-18 in the peripheral blood of Fap fl / fl or Fap ΔGfap mice with MC38-luc or Panc02-luc injection ( n = 5). (G, H) Immunofluorescence staining of NLRP3, caspase-1, or IL-18 (green) in α SMA + HSCs (gray) that adhered to the CD31 + sinusoidal blood vessels (red) in the PMN of CRC or pancreatic cancer ( n = 5). Scale bar, 10 μm. Data are presented as mean ± SEM. ∗ P < 0.05, ∗∗ P < 0.01, ∗∗∗ P < 0.001 (one-way ANOVA with Tukey's post hoc comparison in D; two-tailed unpaired t -test in E–H).

Article Snippet: LX-2 FAP cells were treated with a FAP α inhibitor SP-13786 (50 μmol/L, T7890, TargetMol, Shanghai, China) for 30 min. Gene knockdown in LX-2 Vector and LX-2 FAP cells was achieved by infection with shRNA plasmids or transfection with siRNAs, using corresponding negative controls as references.

Techniques: Analysis, Sequencing, Western Blot, Immunocytochemistry, Enzyme-linked Immunosorbent Assay, Concentration Assay, Injection, Immunofluorescence, Staining, Comparison, Two Tailed Test, Significance Assay