Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: Overlapping of genes resulting from comparison of downregulated genes in LX-2 cells incubated with Exo HCT116 KD and upregulated genes in LX-2 cells incubated with Exo SW480 OE ( A ). Heat map of transcriptome alterations after incubating LX-2 cells with Exo HCT116 KD and Exo HCT116 KD Ctrl ( B ). Heat map of transcriptome alterations after incubating LX-2 cells with Exo SW480 OE and Exo SW480 OE Ctrl ( C ). The relative mRNA level of CXCL5 was analyzed in a group of independent samples ( D ). Western blot showed protein level of CXCL5 in LX-2 cells incubated with Exo HCT116 KD ( E ) and Exo SW480 OE (F) and their relative control respectively. ELISA analysis of CXCL5 level in culture medium from LX-2 cells incubated with Exo HCT116 KD ( G ) and Exo SW480 OE ( H ) and their relative control respectively. Immunofluorescence staining of α-SMA, FAP and CXCL5 in LX-2 cells incubated with Exo HCT116 KD ( I ) and Exo SW480 OE ( J ) and their relative control respectively. Scale bar = 20 μm. Each experiment was performed in triplicate. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001, ns not significant.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Comparison, Incubation, Western Blot, Control, Enzyme-linked Immunosorbent Assay, Immunofluorescence, Staining

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: The effects of acetate treatment on acetyl-CoA levels in LX-2 cells incubated with Exo HCT116 ( A ) and Exo SW480 ( B ). Western blot showed the effects of acetate treatment on CXCL5 and H3K27ac levels in LX-2 cells incubated with Exo HCT116 ( C ) and Exo SW480 ( D ). E The effects of Trichostatin A (TSA) treatment on CXCL5 and H3K27ac levels in LX-2 cells incubated with Exo HCT116 KD and their relative control. F The effects of C646 treatment on CXCL5 and H3K27ac levels in LX-2 cells incubated with Exo SW480 OE and their relative control. G , H Chromatin immunoprecipitation (ChIP) assays using IgG as a control were performed with antibody against H3K23ac. Examination of H3K27 acetylation status in CXCL5 gene promoter region in LX-2 cells incubated with Exo HCT116 KD and Exo HCT116 KD Ctrl or in LX-2 cells incubated with Exo HCT116 KD and Exo HCT116 KD Ctrl and treated with TSA ( G ). Examination of H3K27 acetylation status in CXCL5 gene promoter region in LX-2 cells incubated with Exo SW480 OE and Exo SW480 OE Ctrl or in LX-2 cells incubated with Exo SW480 OE and Exo SW480 OE Ctrl and treated with C646 ( H ). Each experiment was performed in triplicate. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Incubation, Western Blot, Control, Chromatin Immunoprecipitation

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: Western blot showed the protein levels of HSPC111, CXCR2 and EMT-relative genes in CAFs CM-incubated HCT116 ( A ) and SW480 cells ( B ). Migration of HCT116 ( C ) and SW480 cells ( D ) were analyzed by transwell assay after CAFs CM treatment. Scale bar = 200 μm. Western blot showed the protein levels of HSPC111, CXCR2 and EMT-relative genes in CAFs CM- and CXCL5 neutralizing antibody-incubated HCT116 ( E ) and SW480 cells ( F ). G Migration of HCT116 cells were analyzed by wound-healing assay after CAFs CM and CXCL5 neutralizing antibody treatment. Scale bar = 100 μm. Western blot showed the protein levels of HSPC111, CXCR2 and EMT-relative genes in CAFs CM- and CXCR2 inhibitor (navarixin)-incubated HCT116 ( H ) and SW480 cells ( I ). J Migration of HCT116 cells were analyzed by wound-healing assay after CAFs CM and CXCR2 inhibitor (navarixin) treatment. Scale bar = 100 μm. K IVIS imaging on experimental liver metastasis of indicated mice treated with Exo HCT116 KD or their relative control and with/without navarixin. Luciferase-labeled HCT116 cells were used to perform experimental liver metastasis model ( n = 3). L Representative pictures and quantitative results of HE staining of liver tissue sections from indicated mice. Scale bar = 500 μm. Each experiment was performed in triplicate. Data are shown as mean ± SD. * P < 0.05, ** P < 0.01, *** P < 0.001.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Western Blot, Incubation, Migration, Transwell Assay, Wound Healing Assay, Imaging, Control, Luciferase, Labeling, Staining

Journal: Cell Death & Disease

Article Title: Cancer-derived exosomal HSPC111 promotes colorectal cancer liver metastasis by reprogramming lipid metabolism in cancer-associated fibroblasts

doi: 10.1038/s41419-022-04506-4

Figure Lengend Snippet: Exosomal HSPC111 derived from CRC cells phosphorylates ACLY in CAFs, leading to the increased levels of acetyl-CoA and histone acetylation to secrete CXCL5, and resulting in CRC cells colonized in liver via the CXCL5-CXCR2 axis.

Article Snippet: Cells were cultured in the presence of appropriate exosomes (20 μg/mL) for 48 h before supernatants were collected, and the CXCL5 level was detected by CXCL5 ELISA kit (EK0728, BOSTER) according to the manufacturer’s protocols.

Techniques: Derivative Assay