vcap human prostate cancer cell line  (ATCC)

Journal: Advanced Science

Article Title: O‐GalNAc Glycosylation Activates MBL‐Mediated Complement and Coagulation Cascades to Drive Organotropic Metastasis

doi: 10.1002/advs.202504809

Figure Lengend Snippet: High MBL pathway activity is enriched in PCa liver metastasis and GalNAc is a major glycosylation form on NEPC cell surface. a) The KEGG analysis showing that the “complement and coagulation cascades” is most upregulated signature in liver metastasis versus primary prostate tumors of rb1 Δ/Δ p53 Δ/Δ NEPC tumor‐bearing mice (n = 3, mice). b,c) GSEA plots reveal that “complement and coagulation cascade” (b) and “MBL pathway activity” are significantly elevated in liver metastatic lesions compared to metastasis in other anatomic sites based on analysis of the SU2C prostate cancer dataset. d) Kaplan–Meier survival analysis of the SU2C prostate cancer dataset reveal that PCa patients with high MBL activity (n = 39) exhibit significantly shorter survival compared to PCa patients with low MBL activity (n = 39). The log rank‐test was applied for statistics. e,f) GSEA plots showing that the “cell surface glycosylation” is significantly elevated in NEPC compared to PrAD based on analysis of Beltran and SU2C prostate cancer datasets. g) Lectin microarray results that the O‐GalNAc glycosylation, as revealed by the strongest signal of vicia villosa (VVA) lectin toward the membrane fraction extracted from murine rb1 Δ/Δ p53 Δ/Δ NEPC organoids.

Article Snippet: Human prostate cancer (PCa) cell lines used in this study include VCaP, and LASCPC‐01, which were purchased from American Type Culture Collection (ATCC).

Techniques: Activity Assay, Glycoproteomics, Coagulation, Microarray, Membrane

Journal: Advanced Science

Article Title: O‐GalNAc Glycosylation Activates MBL‐Mediated Complement and Coagulation Cascades to Drive Organotropic Metastasis

doi: 10.1002/advs.202504809

Figure Lengend Snippet: Inhibition of O‐GalNAc glycosylation or Galnt9‐KD attenuates MBL binding and activation and inhibits liver metastasis in NEPC. a) The volcano pot depicts the upregulated, downregulated, and unchanged genes, which encode the enzymes involved in the first two steps of glycosylation based on Beltran human prostate cancer dataset. b) Immunoblots showing the protein level of Galnt9 in WT prostate, myc hi pten Δ/Δ PrAD, and rb1 Δ/Δ p53 Δ/Δ NEPC organoids. c) Immunoblots showing the protein level of GALNT9 in VCaP, LAPC4, 22Rv1, Du145, PC3 and LASCPC‐01 cells. d,e) IF staining and median fluorescence intensity (MFI) quantification showing the upregulated O‐GalNAc glycosylation in rb1 Δ/Δ p53 Δ/Δ NEPC organoids compared to myc hi pten Δ/Δ PrAD counterparts (scale bar = 20 µm). f–g) IF staining and MFI quantification showing the upregulated O‐GalNAc glycosylation in rb1 Δ/Δ p53 Δ/Δ NEPC organoids compared to myc hi pten Δ/Δ PrAD counterparts (scale bar = 10 µm). h–j) O‐GalNAc inhibitor Benzyl‐α‐GalNAc (5 µ m , treated for 24 h) significantly suppressed the MBL binding and activation capabilities of rb1 Δ/Δ p53 Δ/Δ NEPC organoids. k,l) IF staining and MFI quantification showing the upregulated O‐GalNAc glycosylation in rb1 Δ/Δ p53 Δ/Δ ‐scramble and rb1 Δ/Δ p53 Δ/Δ ‐shGalnt9 NEPC organoids (scale bar = 50 µm). m–o) Galnt9 ‐KD in rb1 Δ/Δ p53 Δ/Δ NEPC organoids resulted in significantly decreased MBL binding (m‐n) and activation capabilities. p–q) IF staining images and MFI quantification revealed the O‐GalNAc glycosylation status in LAPC4, VCaP, and LASCPC‐01 cells (scale bar = 10 µm). r–t) GALNT9 ‐KD in human NEPC LASCPC‐01 cells resulted in significantly decreased MBL binding and activation capabilities. u–w) Dissected livers, H&E staining images, and quantification of the liver metastatic foci number w) of the C57BL/6 recipients inoculated with rb1 Δ/Δ p53 Δ/Δ ‐scramble (n = 7, mice) and rb1 Δ/Δ p53 Δ/Δ ‐sh Galnt9 organoids (n = 8, mice). For statistics in this figure, the two‐tail unpaired Student's‐ t test was applied for (e), (g), (i‐j) and (w), and the one‐way ANOVA test was applied for (n‐o), (q), and (s‐t). Data were shown as means ± SD.

Article Snippet: Human prostate cancer (PCa) cell lines used in this study include VCaP, and LASCPC‐01, which were purchased from American Type Culture Collection (ATCC).

Techniques: Inhibition, Glycoproteomics, Binding Assay, Activation Assay, Western Blot, Staining, Fluorescence