Journal: Biosensors

Article Title: Malignant Melanoma-Derived Exosomes Induce Endothelial Damage and Glial Activation on a Human BBB Chip Model

doi: 10.3390/bios12020089

Figure Lengend Snippet: Exosomes derived from A375 cells were more easily absorbed by human brain microvascular endothelial cells. ( A ) Schematic diagram of exosomes purification from tumor cells by differential centrifugation. ( B ) Western blotting showing protein level of exosomal markers (CD63, CD81) in 4 types of tumor cell-derived exosomes ( n = 3). ( C ) Representative TEM images of 4 types of tumor cell-derived exosomes ( n = 2). ( D ) Representative confocal images of hCMEC/D3 cells 24 h after incubation with exosomes (2 μg/mL and 10 μg/mL) derived from 4 different types of tumor cells ( n = 4). Exosomes were labeled by PKH67 (green) fluorescent dye. ( E ) Quantification of PKH67 fluorescence intensity for each group based on ( D ) ( n = 4). Data are presented as mean ± SEM, and are analyzed using a one-way ANOVA with Bonferroni post-test (***: p < 0.001).

Article Snippet: A375 cells (iCell, #A375) were cultured in DMEM medium (Gibco, #C11995500BT) supplemented with 10% FBS and 1% P/S.

Techniques: Derivative Assay, Purification, Centrifugation, Western Blot, Incubation, Labeling, Fluorescence

Journal: Biosensors

Article Title: Malignant Melanoma-Derived Exosomes Induce Endothelial Damage and Glial Activation on a Human BBB Chip Model

doi: 10.3390/bios12020089

Figure Lengend Snippet: A375 cell-derived exosomes caused BBB damage and glial activation. ( A ) Side views of the BBB chip 24 h later following exposure of A375 cell-derived exosomes (green) to the vascular side ( n = 3). ( B ) Representative confocal images of glial cells 24 h later following exposure of A375 cell-derived exosomes labeled by PKH67 dye to the vascular side ( n = 3). ( C ) BBB permeability assay of 10 kDa FITC-dextran following exposure of A375 cell-derived exosomes to the vascular side ( n = 4). Data are presented as mean ± SEM, and are analyzed using unpaired Student’s t -test (*: p < 0.05). ( D ) Representative confocal images of brain endothelial cells (VE-cadherin) 72 h later following treatment of A375 cell-derived exosomes ( n = 3). The areas indicated by the white boxes were enlarged on the right. ( E ) Western blotting showing the protein level of VE-cadherin and ZO-1 in brain endothelial cells 72 h later following treatment of A375 cell-derived exosomes ( n = 3). GAPDH was used as an internal control. ( F , G ) Representative confocal images of astrocytes (GFAP; F ) and microglia (IBA1; G ) 72 h later following treatment of A375 cell-derived exosomes ( n = 3). ( H ) Bar graph showing IL-6 concentration in culture supernatant of vascular channel 72 h later following exposure of A375 or A549 cell-derived exosomes to the vascular side detected by ELISA kit ( n = 3). ( I ) Bar graph showing IL-6 concentration in culture supernatant of glial channel 72 h later following exposure of A375 or A549 cell-derived exosomes to the vascular side detected by Elisa kit ( n = 3). ( H , I ) Data are presented as mean ± SEM, and are analyzed using a one-way ANOVA with Bonferroni post-test (**: p < 0.01; ***: p < 0.001). ( J ) Representative confocal images of brain endothelial cells and glial cells stained with PI dye 72 h later following treatment of A375 cell-derived exosomes. Cells positive for PI were indicated by white arrowheads ( n = 4). ( K ) Bar graph showing the percentage of PI + cells for each group based on ( J ) ( n = 4). Data are presented as mean ± SEM, and are analyzed using unpaired Student’s t -test (***: p < 0.001).

Article Snippet: A375 cells (iCell, #A375) were cultured in DMEM medium (Gibco, #C11995500BT) supplemented with 10% FBS and 1% P/S.

Techniques: Derivative Assay, Activation Assay, Labeling, Permeability, Western Blot, Control, Concentration Assay, Enzyme-linked Immunosorbent Assay, Staining

Journal: Biosensors

Article Title: Malignant Melanoma-Derived Exosomes Induce Endothelial Damage and Glial Activation on a Human BBB Chip Model

doi: 10.3390/bios12020089

Figure Lengend Snippet: RNA-seq analysis of brain endothelial cells and glial cells following treatment of A375 cell-derived exosomes on the BBB chip. ( A ) Heat-map showing the transcriptional changes of brain endothelial cells and glial cells on BBB chip 3 days after treatment of A375 cell-derived exosomes ( n = 3). ( B , C ) Volcano plots showed the modulated genes in brain endothelial cells ( B ) or glial cells ( C ) 3 days after treatment of A375 cell-derived exosomes. Genes differentially expressed with fold-change >1.5 and p < 0.05 were marked in color. p -values were calculated using a two-sided, unpaired Student’s t-test with equal variance assumed. ( D ) Venn diagrams depicted the down-regulated DEGs shared or unique between brain endothelial cells and glial cells. ( E ) Venn diagrams depicted the up-regulated DEGs shared or unique between brain endothelial cells and glial cells. ( F , G ) Dotplot showed the enriched GO terms based on up-regulated ( F ) or down-regulated ( G ) genes in brain endothelial cells following treatment of A375 cell-derived exosomes. ( H , I ) Dotplot showed the enriched GO terms based on up-regulated ( H ) or down-regulated ( I ) genes in glial cells following treatment of A375 cell-derived exosomes. ( F – I ) The color of the dots represents the rich factor, and the size represents the input number for each GO term.

Article Snippet: A375 cells (iCell, #A375) were cultured in DMEM medium (Gibco, #C11995500BT) supplemented with 10% FBS and 1% P/S.

Techniques: RNA Sequencing, Derivative Assay

Journal: Genes

Article Title: Integrative Transcriptomic Analysis Identify Potential m6A Pathway-Related Drugs That Inhibit Cancer Cell Proliferation

doi: 10.3390/genes13112011

Figure Lengend Snippet: Effect of R428 on human melanoma cell line cell line A375. ( A , B ) The proliferation of the human breast cancer cell line A375 was significantly inhibited by R428 treatment. The results from the cell count assay ( A ) and the MTT assay ( B ) are shown. Data represented the mean ± SEM from five independent experiments, t -test, * p < 0.05, ** p < 0.01. Some SEM values in the drug-treated experimental group that are too small to be shown are specified here: ( A ) 24 h. 0.009; 48 h, 0.012; 72 h, 0.006; 96 h, 0.000; (B) 24 h. 0.004; 48 h, 0.002; 72 h, 0.006; 96 h, 0.002. ( C , D ) The protein expression of METTL3 in the A375 cell line was significantly reduced by drug treatment, t -test, ** p < 0.01. The summary of Western blot grey values as the mean ± SEM from three independent experiments ( C ) and the representative Western blot result ( D ) are shown. ( E ) The RNA m6A modification level in the A375 cell line was significantly decreased after R428 treatment. Data represented the mean ± SEM from five independent experiments, t -test, * p < 0.05. ( F , G ) Representative images of treated and control cells observed under an inverted 10× microscopy.

Article Snippet: The breast cancer cell line MCF7 and the human melanoma cell line A375 were provided by Biomol Inc. (Hamburg, Germany).

Techniques: Cell Counting, MTT Assay, Expressing, Western Blot, Modification, Control, Microscopy

Journal: Journal of Nanobiotechnology

Article Title: Engineering endoplasmic reticulum targeted metal–polyphenol curcumin nanomicelles for melanoma therapy

doi: 10.1186/s12951-025-03941-7

Figure Lengend Snippet: TCPC inhibits melanoma cell proliferation, migration, and invasion in vitro. ( A ) Cell viability of A375 and SK-MEL-28 cells treated with CTRL, PC, TC, or TCPC for 24 h measured by CCK-8 assay. ( B, C ) Wound healing assays of A375 and SK-MEL-28 cells after treatment. ( D, E ) Representative images and quantification of Transwell invasion assays in both cell lines. Scale bar = 200 μm. Data are shown as mean ± SD ( n = 3). Statistical significance was determined by two-sample t-test comparing each treatment group with CTRL. * p < 0.05, ** p < 0.01

Article Snippet: The A375 and SK-MEL-28 line were obtained from Procell (Procell, China).

Techniques: Migration, In Vitro, CCK-8 Assay

Journal: Journal of Nanobiotechnology

Article Title: Engineering endoplasmic reticulum targeted metal–polyphenol curcumin nanomicelles for melanoma therapy

doi: 10.1186/s12951-025-03941-7

Figure Lengend Snippet: Transcriptomic analysis reveals ER stress induction by TCPC in melanoma cells. ( A ) PCA plot showing clear separation between CTRL and TCPC-treated A375 cells. ( B ) Volcano plot of DEGs. ( C ) Bar chart summarizing upregulated and downregulated DEGs. ( D ) Heatmap of representative DEGs. ( E ) GO enrichment analysis highlighting ER-related terms in the cellular component category. ( F ) GSEA enrichment plots of top pathways after TCPC treatment. ( G ) Enrichment plot for “response to ER stress” pathway. Data are presented as mean ± SD ( n = 5)

Article Snippet: The A375 and SK-MEL-28 line were obtained from Procell (Procell, China).

Techniques:

Journal: Journal of Nanobiotechnology

Article Title: Engineering endoplasmic reticulum targeted metal–polyphenol curcumin nanomicelles for melanoma therapy

doi: 10.1186/s12951-025-03941-7

Figure Lengend Snippet: TCPC induces ER stress–mediated apoptosis through Ca²⁺ overload and mitochondrial dysfunction. ( A ) qPCR analysis of ER stress–related genes (CHOP, HSPA5, TTC23L) in A375 and SK-MEL-28 cells. ( B ) Confocal imaging confirming CHOP protein upregulation. Scale bar = 20 μm. ( C ) TEM images showing ER swelling in TCPC-treated cells. Scale bars: 5 μm and 1 μm. ( D, E ) ER-Tracker staining and flow cytometry quantification of ER expansion. Scale bar = 20 μm. (F, G ) Intracellular Ca²⁺ changes visualized by confocal microscopy. Scale bar = 20 μm. (H ) JC-1 staining showing loss of mitochondrial membrane potential. Scale bar = 20 μm. (I, J) Intracellular ROS levels after TCPC treatment. Scale bar = 20 μm. ( K ) Flow cytometry analysis of apoptosis. Data are shown as mean ± SD ( n = 3). Statistical significance was determined by two-sample t-test comparing with CTRL group. * p < 0.05, ** p < 0.01

Article Snippet: The A375 and SK-MEL-28 line were obtained from Procell (Procell, China).

Techniques: Imaging, Staining, Flow Cytometry, Confocal Microscopy, Membrane

Journal: Journal of Nanobiotechnology

Article Title: Engineering endoplasmic reticulum targeted metal–polyphenol curcumin nanomicelles for melanoma therapy

doi: 10.1186/s12951-025-03941-7

Figure Lengend Snippet: In vivo antitumor efficacy of TCPC in a melanoma xenograft model. ( A ) Schematic of the treatment schedule in A375 tumor–bearing nude mice. ( B, C ) Tumor volume growth curves and tumor weights after treatment. ( D ) H&E staining of tumor tissues showing apoptosis and necrosis. ( E, F, H, I ) IHC staining for cleaved caspase-3 and Ki-67 in tumor sections. ( G, J ) IHC staining showing increased CHOP expression in TCPC-treated tumors. Scale bar = 100 μm. Data are shown as mean ± SD ( n = 5), Statistical significance was determined by two-sample t-test comparing with CTRL group. * p < 0.05, ** p < 0.01

Article Snippet: The A375 and SK-MEL-28 line were obtained from Procell (Procell, China).

Techniques: In Vivo, Staining, Immunohistochemistry, Expressing