ABSTRACT
Objective:To investigate the mechanisms of TWS119 induced CCR5 expression in hunman γδT cells. Methods:After treatment with various concentrations of TWS119 for 48h, the expression of CCR5 in γδT cells were detected by flow cytometry. The p-STAT3 and GAPDH expression were examined by Western blot analysis. Results: TWS119 could upregulate the expression of CCR5 in dose dependent manner. Western blot analysis revealed that TWS119 inhibit phosphorylation of STAT3,but had no significant impact on GAPDH. In addition, pretreatment of γδT cells with 0. 5 μmol/L STAT3 specific phosphorylation inhibitor Stattic could upregulate the expression of CCR5 and enhance the TWS119 induced CCR5 expression. Conclusion: TWS119 could upregulate CCR5 expression of γδT cells by inhibiting STAT3 phosphorylation in vitro.
ABSTRACT
OBJECTIVE: To study the antitumor activity of TWS119 and its effect on the killing activity of γδT cells in vitro. METHODS: CCK-8 assay was used for detecting the influence of TWS119 on the proliferation of SGC-7901 and γδT cells and the cytotoxic activity of γδT cells to SGC-7901 cells. The cell apoptosis was measured by flow cytometric analysis. The protein expression was examined by Western blot analysis. RESULTS: It was found that TWS119 could inhibit the growth and induce the apoptosis of SGC-7901 cells. The expression of Bcl-2 in SGC-7901 cells treated with TWS119 was downregulated, while p-ERK1/2 and p-STAT3 were up-regulated. In addition, TWS119 at 0.5-4.0μmol · L-1 could promote the growth of γδT cells and enhance the cytotoxic activity of γδT cells to SGC-7901 cells especially at 4.0 μmol · L-1 for 72 h. Furthermore, TWS119 could upregulate the expression of Bcl-2 and inhibit the expression of p-ERK1/2 and p-STAT3. CONCLUSION: TWS119 in some concentrations can inhibit the growth of SGC-7901 cells, promote the 78T cells growth and enhance the cytotoxic activity of γδT cells to SGC-7901 cells, and the mechanism may be involve in Wnt, ERK1/2, Bcl-2 and p-STAT3 signaling pathways.
ABSTRACT
Objective:To investigate the effect of glycogen synthase kinase-3β inhibitor TWS119 on hunman γδT cells growth and phenotypic characteristics. Methods:Using γδT medium to cultivate human peripheral blood γδT cells in vitro. After co-cultured with different concentrations of glycogen synthase kinase-3β( GSK-3β) inhibitor 4, 6-disubstituted pyrrolopyrimidine ( TWS119 ) for indicated time,growth curve and Wnt/β-catenin activation of in each group were determined by CCK-8 and Western blot assays. The CD62L and CD45RA expression theγδT cells were detected using flow cytometry. Results:Wnt/β-catenin pathway ofγδT cells could activate by TWS119. In the first group,TWS119 could upregulate the expression of CD62L and CD45RA in dose dependent manner while inhibit the growth and ratio of γδT cells. In the second group,TWS119 could promote the growth and ratio of γδT cells with the increase in concentration from 0 μmol/L to 4. 0 μmol/L and decreased thereafter. Besides,TWS119 could promote the expression of CD62L in a dose-dependent and had no effect on the CD45RA. Conclusion: Human γδT cells isolated from peripheral blood treated with TWS119 gave rise to two subsets of CD45RA+CD62L+γδT cells and CD62L+γδT cells.