OBJECTIVE: To investigate the effect of Wnt/β-catenin pathway activation by glycogen synthase kinase-3β inhibitor 4,6-disubstituted pyrrolopyrimidine (TWS119) on proliferation and phenotypic characteristics of human nature killer (NK) cells. METHODS: The peripheral blood mononuclear cells (PBMCs) were obtained from healthy volunteers and added to the complete medium containing recombinant human interleukin-2 (rhIL-2) and human AB serum to isolate NK cells from PBMCs. After co-cultured with 0-8.0 μmol/L TWS119 for 72 hours, growth curve and Wnt/β-catenin activation of NK cells in each group were determined by CCK-8 and Western blotting. The CD107a and CD62L (L-selectin) expressions in the NK cells were detected using flow cytometry. RESULTS: NK cells were amplified to (61.76 ± 3.74)% after human PBMCs were cultured for 10 days. The 0-2.0 μmol/L TWS119 could promote the growth of NK cells in a dose-dependent manner, and the proliferation rate gradually dropped when TWS119 was more than 2.0 μmol/L. 0-8.0 μmol/L TWS119 could activate Wnt/β-catenin pathway and up-regulate the expression of CD62L in a dose-dependent manner, but it decreased the expression of CD107a. CONCLUSION: Human NK cells isolated from peripheral blood treated with TWS119 gave rise to early mature CD62L⁺ NK cells.
OBJECTIVE: To investigate the effect of Wnt/β-catenin pathway activation by glycogen synthase kinase-3β inhibitor 4,6-disubstituted pyrrolopyrimidine (TWS119) on proliferation and phenotypic characteristics of human nature killer (NK) cells. METHODS: The peripheral blood mononuclear cells (PBMCs) were obtained from healthy volunteers and added to the complete medium containing recombinant humaninterleukin-2 (rhIL-2) and human AB serum to isolate NK cells from PBMCs. After co-cultured with 0-8.0 μmol/L TWS119 for 72 hours, growth curve and Wnt/β-catenin activation of NK cells in each group were determined by CCK-8 and Western blotting. The CD107a and CD62L (L-selectin) expressions in the NK cells were detected using flow cytometry. RESULTS: NK cells were amplified to (61.76 ± 3.74)% after human PBMCs were cultured for 10 days. The 0-2.0 μmol/L TWS119 could promote the growth of NK cells in a dose-dependent manner, and the proliferation rate gradually dropped when TWS119 was more than 2.0 μmol/L. 0-8.0 μmol/L TWS119 could activate Wnt/β-catenin pathway and up-regulate the expression of CD62L in a dose-dependent manner, but it decreased the expression of CD107a. CONCLUSION:Human NK cells isolated from peripheral blood treated with TWS119 gave rise to early mature CD62L⁺ NK cells.