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I-branching N-acetylglucosaminyltransferase regulates prostate cancer invasiveness by enhancing α5β1 integrin signaling.

Mikami J, Tobisawa Y, Yoneyama T, Hatakeyama S, Mori K, Hashimoto Y, Koie T, Ohyama C, Fukuda M - Cancer Sci. (2016)

Bottom Line: GCNT2-positive cells were significantly lesser in organ-confined disease than in that with extra-capsular extensions, and GCNT2-negative tumors were associated with significantly better prostate-specific antigen-free survival compared with GCNT2-positive tumors.Subsequent functional studies revealed that knockdown of GCNT2 expression in PCa cell lines significantly inhibited cell migration and invasion.In conclusion, GCNT2 expression is closely associated with invasive potential of PCa.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki, Japan.

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Glycolipids carrying I‐antigens play important roles in integrin‐mediated protein kinase B (AKT) phosphorylation and migration of prostate cancer cells. (a) Functional blocking antibodies against α5 and β1 integrins significantly inhibited cell migration in DU145NC and DU145GCNT2KD4 cells. (b) AKT phosphorylation at serine 473 (p‐S473) was inhibited by the AKT inhibitor VIII (AKTi; 10 mM), and cell migration was inhibited in DU145NC cells but not DU145GCNT2KD4 cells. (c) DU145NC cells were cultured with benzyl‐α‐GalNAc (BAG), DL‐threo‐1‐Phenyl‐2‐palmitoylamino‐3‐morpholino‐1‐propanol hydrochloride (PPMP), or DMSO for 48 h. (c) DU145NC cells were cultured with DMSO or BAG on fibronectin (FN)‐coated dishes for 20 min. Depletion of O‐glycan had no effect on AKT p‐S473. (d) DU145NC cells treated with BAG had significantly inhibited cell migration. Depletion of glycolipids in DU145NC cells significantly inhibited AKT p‐S473 (e) and migration (f). Migration assays were carried out in triplicate. NS, not significant; t‐AKT, total‐AKT.
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cas12859-fig-0005: Glycolipids carrying I‐antigens play important roles in integrin‐mediated protein kinase B (AKT) phosphorylation and migration of prostate cancer cells. (a) Functional blocking antibodies against α5 and β1 integrins significantly inhibited cell migration in DU145NC and DU145GCNT2KD4 cells. (b) AKT phosphorylation at serine 473 (p‐S473) was inhibited by the AKT inhibitor VIII (AKTi; 10 mM), and cell migration was inhibited in DU145NC cells but not DU145GCNT2KD4 cells. (c) DU145NC cells were cultured with benzyl‐α‐GalNAc (BAG), DL‐threo‐1‐Phenyl‐2‐palmitoylamino‐3‐morpholino‐1‐propanol hydrochloride (PPMP), or DMSO for 48 h. (c) DU145NC cells were cultured with DMSO or BAG on fibronectin (FN)‐coated dishes for 20 min. Depletion of O‐glycan had no effect on AKT p‐S473. (d) DU145NC cells treated with BAG had significantly inhibited cell migration. Depletion of glycolipids in DU145NC cells significantly inhibited AKT p‐S473 (e) and migration (f). Migration assays were carried out in triplicate. NS, not significant; t‐AKT, total‐AKT.

Mentions: To demonstrate the roles of I‐antigens, cell migration assays were carried out after treatment of DU145 cells with various inhibitors. In these experiments, function‐blocking antibodies against α5 integrin and β1 integrin strongly inhibited DU145NC and DU145GCNT2KD4 cell migration (Fig. 5a). Moreover, the AKT phosphorylation inhibitor had high efficacy in DU145 cells without causing cytotoxicity (Fig. S5a,b), although numbers of migrant cells were significantly fewer among DU145NC cells in which AKT is strongly activated than in DU145GCNT2KD4 cells that express AKT p‐S473 at low levels (Fig. 5b). Because I‐antigens are carried on O‐glycan and glycolipids, we determined which glycan is more important for AKT p‐S473 and cell migration. Although BAG‐treated DU145NC cells had significantly reduced migration potential (Fig. 5d), AKT p‐S473 did not differ between vehicle control and BAG‐treated cells (Fig. 5c). However, after PPMP treatments, DU145NC cells showed strongly inhibited AKT p‐S473 and cell migration (Fig. 5e,f). In addition, cell viability was >90% after BAG and PPMP treatments (Fig. S5c,d), indicating that O‐glycans carrying I‐antigens support cell migration, and that glycolipids carrying I‐antigens play important roles in integrin–fibronectin‐mediated PI3K/AKT activation and cell migration.


I-branching N-acetylglucosaminyltransferase regulates prostate cancer invasiveness by enhancing α5β1 integrin signaling.

Mikami J, Tobisawa Y, Yoneyama T, Hatakeyama S, Mori K, Hashimoto Y, Koie T, Ohyama C, Fukuda M - Cancer Sci. (2016)

Glycolipids carrying I‐antigens play important roles in integrin‐mediated protein kinase B (AKT) phosphorylation and migration of prostate cancer cells. (a) Functional blocking antibodies against α5 and β1 integrins significantly inhibited cell migration in DU145NC and DU145GCNT2KD4 cells. (b) AKT phosphorylation at serine 473 (p‐S473) was inhibited by the AKT inhibitor VIII (AKTi; 10 mM), and cell migration was inhibited in DU145NC cells but not DU145GCNT2KD4 cells. (c) DU145NC cells were cultured with benzyl‐α‐GalNAc (BAG), DL‐threo‐1‐Phenyl‐2‐palmitoylamino‐3‐morpholino‐1‐propanol hydrochloride (PPMP), or DMSO for 48 h. (c) DU145NC cells were cultured with DMSO or BAG on fibronectin (FN)‐coated dishes for 20 min. Depletion of O‐glycan had no effect on AKT p‐S473. (d) DU145NC cells treated with BAG had significantly inhibited cell migration. Depletion of glycolipids in DU145NC cells significantly inhibited AKT p‐S473 (e) and migration (f). Migration assays were carried out in triplicate. NS, not significant; t‐AKT, total‐AKT.
© Copyright Policy - creativeCommonsBy-nc
Related In: Results  -  Collection

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Show All Figures
getmorefigures.php?uid=PMC4814258&req=5

cas12859-fig-0005: Glycolipids carrying I‐antigens play important roles in integrin‐mediated protein kinase B (AKT) phosphorylation and migration of prostate cancer cells. (a) Functional blocking antibodies against α5 and β1 integrins significantly inhibited cell migration in DU145NC and DU145GCNT2KD4 cells. (b) AKT phosphorylation at serine 473 (p‐S473) was inhibited by the AKT inhibitor VIII (AKTi; 10 mM), and cell migration was inhibited in DU145NC cells but not DU145GCNT2KD4 cells. (c) DU145NC cells were cultured with benzyl‐α‐GalNAc (BAG), DL‐threo‐1‐Phenyl‐2‐palmitoylamino‐3‐morpholino‐1‐propanol hydrochloride (PPMP), or DMSO for 48 h. (c) DU145NC cells were cultured with DMSO or BAG on fibronectin (FN)‐coated dishes for 20 min. Depletion of O‐glycan had no effect on AKT p‐S473. (d) DU145NC cells treated with BAG had significantly inhibited cell migration. Depletion of glycolipids in DU145NC cells significantly inhibited AKT p‐S473 (e) and migration (f). Migration assays were carried out in triplicate. NS, not significant; t‐AKT, total‐AKT.
Mentions: To demonstrate the roles of I‐antigens, cell migration assays were carried out after treatment of DU145 cells with various inhibitors. In these experiments, function‐blocking antibodies against α5 integrin and β1 integrin strongly inhibited DU145NC and DU145GCNT2KD4 cell migration (Fig. 5a). Moreover, the AKT phosphorylation inhibitor had high efficacy in DU145 cells without causing cytotoxicity (Fig. S5a,b), although numbers of migrant cells were significantly fewer among DU145NC cells in which AKT is strongly activated than in DU145GCNT2KD4 cells that express AKT p‐S473 at low levels (Fig. 5b). Because I‐antigens are carried on O‐glycan and glycolipids, we determined which glycan is more important for AKT p‐S473 and cell migration. Although BAG‐treated DU145NC cells had significantly reduced migration potential (Fig. 5d), AKT p‐S473 did not differ between vehicle control and BAG‐treated cells (Fig. 5c). However, after PPMP treatments, DU145NC cells showed strongly inhibited AKT p‐S473 and cell migration (Fig. 5e,f). In addition, cell viability was >90% after BAG and PPMP treatments (Fig. S5c,d), indicating that O‐glycans carrying I‐antigens support cell migration, and that glycolipids carrying I‐antigens play important roles in integrin–fibronectin‐mediated PI3K/AKT activation and cell migration.

Bottom Line: GCNT2-positive cells were significantly lesser in organ-confined disease than in that with extra-capsular extensions, and GCNT2-negative tumors were associated with significantly better prostate-specific antigen-free survival compared with GCNT2-positive tumors.Subsequent functional studies revealed that knockdown of GCNT2 expression in PCa cell lines significantly inhibited cell migration and invasion.In conclusion, GCNT2 expression is closely associated with invasive potential of PCa.

View Article: PubMed Central - PubMed

Affiliation: Department of Urology, Hirosaki University Graduate School of Medicine, Hirosaki, Japan.

Show MeSH
Related in: MedlinePlus