Ligand Discovery for the Alanine-Serine-Cysteine Transporter (ASCT2, SLC1A5) from Homology Modeling and Virtual Screening
Fig 8
Chloroalanine, AOC and γ-FBP inhibit ASCT2-mediated glutamine uptake and cell viability in C8161 human melanoma cells.
(A-C) [3H]-L-glutamine uptake in C8161 cells was used to determine the IC50 of chloroalanine, AOC and γ-FBP (n = 3). (D-F) MTT cell viability assay (n = 3) in C8161 cells incubated with chloroalanine (5 mM), AOC (5 mM) and γ-FBP (5 mM). Two-way ANOVA test was performed to determine significance. (G-I) Apoptosis (Early, Annexin V+ PI-; Late, Annexin V+PI+) was examined by flow cytometry in C8161 cells incubated with chloroalanine (5 mM), AOC (5 mM) and γ-FBP (5 mM). Mann Whitney U test was used to determine significance. (J) ASCT2 expression (with GAPDH as a loading control) in C8161 cells was assessed after 48 hours incubation with chloroalanine (5 mM), AOC (5 mM) and γ-FBP (5 mM).