Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: ( A ) The hypothesis: Co-culture with M2 macrophages or inducing with M2-Exo induces EMT in HT-29 cells. ( B ) Invert phase-contrast images of HT-29 cells before and after co-culture with M2 macrophages. ( C ) Cell viability assay images showing the biocompatibility of co-culture conditions at 1:2, 1:1, and 2:1 cell ratio of HT-29 to M2 macrophages. ( D ) Immunofluorescent microscopy images obtained for E-Cad (green) and N-Cad (red), two EMT markers, obtained with HT-29 cells before and after co-culture with M2 macrophages. ( E ) Gene expression analyses for EMT (E-Cad and N-Cad) and metastasis (MMP2 and MMP9) markers at varying HT-29:M2 macrophage cell number ratio (1:2, 1:1, 2:1). All conditions for gene expression analysis were performed in triplicate using independent Transwell setups. All schematic illustrations were created by the authors using BioRender.

Article Snippet: The comprehensive analysis conducted in this study elucidated the molecular signatures associated with M2-Exosome-mediated post-tumorigenic mechanisms in HT-29 cells.

Techniques: Co-Culture Assay, Viability Assay, Microscopy, Gene Expression

Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: ( A ) Invert phase-contrast images showing the mesenchymal transiting cells upon M2-Exo induction to HT-29 cells with varying initial cell number (50 K, 100 K, and 200 K). ( B ) An illustration depicting the observations in microscopical assessments. ( C ) Cell proliferation test showing the proliferative ability of HT-29 cells against M2-Exo induction. ( D ) Cell viability assay testing the biocompatibility of M2-Exo induction. ( E ) EMT (E-Cad and N-Cad) gene expressions in HT-29 cells treated with M2-Exo. For cell proliferation, each red dot represents a single biological replicate. All RT-qPCR experiments were performed across three independent biological replicates. All schematic illustrations were created by the authors using BioRender.

Article Snippet: The comprehensive analysis conducted in this study elucidated the molecular signatures associated with M2-Exosome-mediated post-tumorigenic mechanisms in HT-29 cells.

Techniques: Viability Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: ( A ) Invert phase-contrast images showing the mesenchymal transiting cells upon M2-Exo induction with varying M2-Exo concentration (50, 100, and 200 μg.mL -1 ). ( B ) Cell viability assay testing the biocompatibility of low, mid, and high doses of M2-Exo. ( C ) Cell proliferation test showing the proliferative ability of HT-29 cells against varying doses of M2-Exo. ( D ) EMT (E-Cad and N-Cad) and metastatic (MMP-2 and MMP-9) gene expressions in HT-29 cells treated with varying doses of M2-Exo. ( E ) Immunofluorescent images were obtained with E-Cad and N-Cad before and after M2-Exo treatment of HT-29 cells. For cell proliferation, each red dot represents a single biological replicate. All RT-qPCR experiments were performed in triplicate across three independent biological replicates.

Article Snippet: The comprehensive analysis conducted in this study elucidated the molecular signatures associated with M2-Exosome-mediated post-tumorigenic mechanisms in HT-29 cells.

Techniques: Concentration Assay, Viability Assay, Quantitative RT-PCR

Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: Effect of exosome loading frequency on EMT. ( A ) The rationality of loading frequency optimization. ( B ) Cell proliferation ability of HT-29 cells in response to different M2-Exo loading frequency. ( C ) Cell viability test for HT-29 cells treated with M2-Exo with different loading frequency. ( D ) Assessment of EMT in HT-29 cells after M2-Exo exposure for 1-, 2-, and 3 times. For cell proliferation, each red dot represents a single biological replicate. All RT-qPCR experiments were performed in triplicate across three independent biological replicates. All schematic illustrations were created by the authors using BioRender.

Article Snippet: The comprehensive analysis conducted in this study elucidated the molecular signatures associated with M2-Exosome-mediated post-tumorigenic mechanisms in HT-29 cells.

Techniques: Quantitative RT-PCR

Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: Transcriptomic analysis to elucidate the mechanism of M2-Exo–initiated EMT. ( A ) Verification of M2-Exo mediated upregulation of EMT in HT-29 cells preceding transcriptomic analysis. ( B ) PCA plot of RNA-seq data illustrating transcriptomic distinctions in HT-29 cells co-cultured with M2 macrophages and stimulated with M2-Exo, with HT-29 cells alone as the negative control. ( C ) A Venn diagram illustrating common and discriminated transcript alterations across groups. ( D ) Hierarchical clustering analysis of transcriptomic alterations reveals distinct patterns in HT-29 cells stimulated with M2-Exo compared to both co-culture and HT-29-only groups. Volcano scatter plots illustrating differential up/down-regulation of transcripts among groups: ( E ) co-cultured vs HT-29 cells alone, ( F ) M2-Exo–stimulated HT-29 cells vs HT-29 cells alone, and ( G ) HT-29 cells co-cultured with M2 macrophages and stimulated with M2-Exo. ( H ) KEGG pathway , analysis identified gene expression changes in M2-Exo–treated HT-29 cells, consistent with cancer-related pathways. ( I ) Identification of EMT-related upregulated transcripts in M2-Exo stimulated HT-29 cells through transcriptome analysis, followed by RT-qPCR validation demonstrating specific upregulation mediated by M2-Exo, distinct from co-culture and HT-29-only groups. All transcriptomic analyses were performed using two biological replicates, while RT-qPCR experiments were conducted in the biological replicates.

Article Snippet: The comprehensive analysis conducted in this study elucidated the molecular signatures associated with M2-Exosome-mediated post-tumorigenic mechanisms in HT-29 cells.

Techniques: RNA Sequencing, Cell Culture, Negative Control, Co-Culture Assay, Gene Expression, Quantitative RT-PCR, Biomarker Discovery

Journal: Scientific Reports

Article Title: M2 macrophage–derived extracellular vesicles induce EMT-like transcriptional reprogramming in colorectal cancer cells via upregulation of FAM83A

doi: 10.1038/s41598-026-39262-4

Figure Lengend Snippet: Validation of M2-Exo stimulated FAM83A-mediated EMT activation through FAM83A silencing and examining by RT-qPCR analysis. ( A ) Silencing of FAM83A in M2-Exo–induced HT-29 cells using varying concentrations of FAM83A-siRNA (sir10, 20, 50 nM) and corresponding scramble-siRNA controls (scr10, 20, 50 nM). HT-29 cells alone and HT-29 cells stimulated with M2-Exo served as negative and positive controls, respectively. Silencing FAM83A attenuates M2-Exo–mediated EMT, as demonstrated by reduced expression of the ( B ) mesenchymal marker N-Cadherin and ( C ) the metastasis marker MMP-2, in M2-Exo stimulated HT-29 cells, respectively. ( D ) FAM83A expression was assessed in breast cancer (MCF-7), melanoma (451LU), and glioblastoma (T98G) cells without M2-Exosome stimulation (NS = p > 0.05, *p < 0.05, **p < 0.01, ***p < 0.001). Western Blot images obtained with N-cadherin and E-cadherin to assess the response of HT-29 cells against M2-Exo, M2-Exo–Akt inhibitor co-load, M2-Exo–siRNA co-load, and M2-Exo–Akt inhibitor-siRNA co-load. RT-qPCR experiments were conducted in triplicate across three biological replicates. Western blot analysis was performed in duplicate using protein samples pooled from three independent biological replicates.

Article Snippet: The comprehensive analysis conducted in this study elucidated the molecular signatures associated with M2-Exosome-mediated post-tumorigenic mechanisms in HT-29 cells.

Techniques: Biomarker Discovery, Activation Assay, Quantitative RT-PCR, Expressing, Marker, Western Blot

Journal: Discover Oncology

Article Title: Functional role of cancer stem cell like exosomes on survival and drug resistance behaviors of colorectal cancer cells

doi: 10.1007/s12672-025-04295-0

Figure Lengend Snippet: CSCs-, HT-29- and Caco-2-derived exosomes decreased the proliferation of Caco-2 and HT-29 cells. A Comparative analysis of Caco-2 cell proliferation after treatment with exosomes at different time intervals (24, 48, and 72 h) ( n = 3). As observed in the graph, exosome-treated groups (CSCs-, HT-29-, and Caco-2-derived exosomes) showed decreased cell proliferation compared to untreated groups and PBS. Cell proliferation was higher in cells treated with CSCs- derived exosomes compared to cells treated with HT-29-, and Caco-2-derived exosomes. B Comparative analysis of HT-29 cell proliferation after treatment with exosomes at 48 h ( n = 3). Exosome-treated groups showed a significant decrease in cell proliferation compared to untreated and PBS-treated groups. Following Bonferroni correction, the difference in cell proliferation was statistically significant ( p < 0.05) between CSCs-EXOs and HT-29-EXOs treated groups. The p-values less than 0.05, 0.01, 0.001, and 0.0001 are shown using *, **, ***, and ****, respectively, on the graphs ( n = 3). C Comparative analysis of cell proliferation following 48 h treatment with CSC-derived exosomes (CSCs-EXOs), heat-inactivated exosomes (HI-EXOs; 95 °C for 10 min), or PBS as a control. Significant reductions in proliferation were observed in cells treated with CSC-EXOs, whereas HI-EXOs failed to induce this effect and showed proliferation levels comparable to the control (p-value < 0.01). These results indicate that the antiproliferative response depends on the biological activity and functional integrity of exosomal biomolecules. Data represent mean ± SD from three independent experiments; p < 0.05 versus the control

Article Snippet: B Comparative analysis showed that Caco-2 cell proliferation was significantly reduced after 5-FU treatment in exosome-treated groups (CSCs-, Caco-2- and HT-29-EXOs) compared to the control groups.

Techniques: Derivative Assay, Control, Activity Assay, Functional Assay

Journal: Discover Oncology

Article Title: Functional role of cancer stem cell like exosomes on survival and drug resistance behaviors of colorectal cancer cells

doi: 10.1007/s12672-025-04295-0

Figure Lengend Snippet: The cellular viability of Caco-2 cells treated with 5-FU chemotherapy drug and the effect of exosomes on drug cytotoxicity properties of Caco-2 cells. A IC50 and R2 square for different time points of 5-FU (24, 48, and 72 h) were 11.52 (R2 = 0.8148), 10.09 (R2 = 0.9020), and 6.4941 (R2 = 0.9130), respectively. Cellular viability was quantified by MTT assay. Data were recorded as the mean ± SD. B Comparative analysis showed that Caco-2 cell proliferation was significantly reduced after 5-FU treatment in exosome-treated groups (CSCs-, Caco-2- and HT-29-EXOs) compared to the control groups. While, cell proliferation was higher in cells treated with 5-FU + H29 CSCs-EXOs compared to cells treated with 5-FU + Caco-2-Exos and 5-FU + Caco-2-EXOs, but it was not statistically significant. P-values less than 0.05, 0.01, 0.001 and 0.0001 were shown using *, **, ***and ****, respectively, on the graphs ( n = 3)

Article Snippet: B Comparative analysis showed that Caco-2 cell proliferation was significantly reduced after 5-FU treatment in exosome-treated groups (CSCs-, Caco-2- and HT-29-EXOs) compared to the control groups.

Techniques: MTT Assay, Control