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: 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 promotes breast cancer cell proliferation. a Crystal violet staining of BT-549 and Hs578T cells treated with DPBS or BCAF-derived exosomes (left). Quantification of cell numbers (middle). qRT‒PCR analysis of miR-1290 expression in recipient cells (right). b Crystal violet staining of BT-549 and Hs578T cells treated with DPBS- or MB-231-derived exosomes (left). Quantification of cell numbers (middle). qRT‒PCR analysis of miR-1290 expression in recipient cells (right). Scale bar, 500 μm. c Crystal violet staining of BT-549 and Hs578T cells transfected with the miR-1290 mimic or mimic-NC (left). Quantification of cell numbers (middle). qRT‒PCR analysis of miR-1290 expression (right). Scale bar, 500 μm. d Representative images of xenograft tumors derived from BT-549 cells transfected with the miR-1290 mimic or mimic-NC. e Growth curves of xenograft tumors in NOD/Shi-scid/IL-2Rγ null mice inoculated with BT-549 cells transfected with the miR-1290 mimic or mimic-NC ( n = 6 per group). The tumor volume was compared between the two groups at the experimental endpoint. f Tumor weight measurements at the endpoint of the xenograft experiment shown in ( e ) ( n = 6 per group). g Representative immunocytochemistry images showing the localization and expression of Ki67 in BT-549 and Hs578T cells treated with BCAF exosomes or MB-231 exosomes and transfected with the miR-1290 mimic or mimic-NC. The nuclei were counterstained with DAPI. Scale bar, 150 μm. The data are shown as the means ± SDs of three independent experiments unless otherwise indicated, and statistical significance was determined using Student’s t tests (* p < 0.05, ** p < 0.01, *** p < 0.001)

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: Staining, Derivative Assay, Exosomes, Analysis, Expressing, Transfection, Xenograft Assay, Immunocytochemistry

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 promotes breast cancer cell proliferation by directly targeting TBC1D4. a Venn diagram analysis of genes downregulated according to the RNA-seq data of MB-231 exosome-treated (FPKM ratio < 0.7 versus DPBS; n = 981) and miR-1290 mimic-transfected BT-549 cells (FPKM ratio < 0.5 versus mimic-NC; n = 676) with predicted miR-1290 targets from TargetScan ( n = 4601), identifying 12 common candidate genes (left). Expression of TBC1D4 under the indicated conditions (right). b Expression levels of TBC1D4 in normal ( n = 114) and breast cancer ( n = 1097) tissues from the TCGA database (*** p < 0.001, Mann‒Whitney U test). c Kaplan‒Meier plot of the overall survival of breast cancer patients from the TCGA database. The survival rate of the high-TBC1D4 subgroup was significantly greater than that of the low-TBC1D4 subgroup ( p = 0.003, log-rank test). d qRT‒PCR analysis of TBC1D4 expression in BT-549 and Hs578T cells treated with the indicated conditions. e Western blot analysis of TBC1D4 expression in BT-549 and Hs578T cells under the indicated conditions. ACTB was used as a loading control. f Representative immunocytochemistry images showing the localization and expression of TBC1D4 in BT-549 and Hs578T cells treated with BCAF exosomes and transfected with the miR-1290 mimic or mimic-NC. The nuclei were counterstained with DAPI. Scale bar, 150 μm. g Dual-luciferase reporter assay in HEK293 cells cotransfected with wild-type or mutant TBC1D4 3’-UTR luciferase reporter vectors and miR-1290 mimic or mimic-NC (six technical replicates) (n.s. not significant). h qRT‒PCR analysis of TBC1D4 expression in BT-549 and Hs578T cells transfected with siTBC1D4 or siCont. i Western blot analysis of TBC1D4 expression in BT-549 and Hs578T cells transfected with siTBC1D4 or siCont. ACTB was used as a loading control. j Crystal violet staining of BT-549 and Hs578T cells transfected with siTBC1D4 or siCont and quantification of cell numbers. Scale bar, 500 μm. k Representative immunocytochemistry images showing the localization and expression of Ki67 in BT-549 and Hs578T cells transfected with siTBC1D4 or siCont. The nuclei were counterstained with DAPI. Scale bar, 150 μm. The data are shown as the means ± SDs of three independent experiments, and statistical significance was determined using Student’s t tests (*** p < 0.001)

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: Analysis, RNA Sequencing, Transfection, Expressing, Western Blot, Control, Immunocytochemistry, Exosomes, Luciferase, Mutagenesis, Staining

Journal: International Journal of Molecular Sciences

Article Title: Proteasome-Enriched hPPSCs-Derived EVs Attenuate Hypoxic Injury in Endothelial Cells via Proteasome-Mediated HIF-1α Degradation

doi: 10.3390/ijms27157013

Figure Lengend Snippet: Characterization and proteomics of human placenta-derived perivascular stem cells (hPPSCs)–extracellular vesicles (EVs). ( A ) Transmission electron microscopy (TEM) characterization of EVs. Scale bar = 200 nm. The arrow is pointing at the EV. ( B ) Nanoparticle tracking analysis (NTA) of EVs. ( C ) WB analysis of ALIX, CD63, and CD81 in EVs. Equal total proteins were loaded per lane for EV samples from two stem cell types. ( D ) Quantitative analysis of ALIX, CD63, and CD81 protein levels. ( E ) WB analysis of Calnexin in EVs and hPPSCs. ( F ) Volcano plot analysis of differentially expressed proteins (DEPs) between hUCMSCs-EVs and hPPSCs-EVs. Green, red, and blue dots represent downregulated proteins, upregulated proteins, and proteins with no significant changes, respectively. The critical criteria were set to FC > 1.2 or FC < 0.83 (corresponding to |log 2 FC| > 0.263), with p < 0.05. ( G ) Protein domain analysis. ( H , I ) GO functional annotation biological process (BP) and GO functional annotation cell composition (CC) enrichment bubble analysis. The closer to red, the smaller the p -value. ( J ) KEGG enrichment analysis. The abscissa denotes the ratio, while the ordinate stands for the category. n = 3, p < 0.05.

Article Snippet: The antibodies employed for Western blot in this study are listed below: Anti-ALIX antibody (Abcam, ab275377, rabbit, Cambridge, MA, USA), diluted at 1:1000; Anti-CD81 antibody (ABclonal, A5270, rabbit, Woburn, MA, USA), diluted at 1:1000; Anti-CD63 antibody (ABclonal, A19023, rabbit), diluted at 1:1000; Anti-HIF-1α antibody (ABclonal, A16873, rabbit), diluted at 1:1000; Anti-Ubiquitin (P4D1) antibody (SANTA, sc-8017, mouse), diluted at 1:1000; Anti-PSMD11 antibody (ABclonal, A15306, rabbit), diluted at 1:500; Anti-PSMB5 antibody (ABclonal, A1975, rabbit), diluted at 1:1000; Anti-Proteasome 20S alpha + beta antibody (Abcam, ab22673, rabbit), diluted at 1:1000; Anti-GAPDH antibody (ABclonal, A19056, rabbit), diluted at 1:5000; Anti-Calnexin antibody (proteintech, 10427-2-AP, rabbit), diluted at 1:10,000; HRP-linked anti-rabbit IgG (CST, 7074, goat), diluted at 1:10,000; and HRP-linked anti-mouse IgG (CST, 7076, horse), diluted at 1:10,000.

Techniques: Derivative Assay, Transmission Assay, Electron Microscopy, Functional Assay

Journal: Medicine

Article Title: Human amniotic epithelial cell-derived exosomes promote conjunctival goblet cells proliferation and mucin secretion

doi: 10.1097/MD.0000000000049647

Figure Lengend Snippet: Identification and characterization of hAECs-Exo. (A) Morphology of exosomes in the hAECs-Exo group was observed under transmission electron microscopy (Scalar bar = 200 nm). (B) Diameter of exosomes in the hAECs-Exo group was analysis under NTA. (C) Western blot was used to observe the expression of CD63, CD81, and Grp94 proteins in hAECs-Exo. hAECs-Exo = human amniotic epithelial cell-derived exosomes, NTA = nanoparticle tracking analysis.

Article Snippet: The polyvinylidene fluoride membrane was treated with protein-free rapid blocking solution (PS108P, EpiZyme) for 15 minutes, and then incubated with primary antibodies against exosome markers CD63 (1:1000, ABclonal, A19023) and CD81 (1:1000, ABclonal, A4863), as well as the negative marker Grp94 (1:1000, ABclonal, A0989) overnight.

Techniques: Transmission Assay, Electron Microscopy, Western Blot, Expressing, Derivative Assay