Effect of cancer-testis antigen OY-TES-1 expression on the polarization of M2 macrophages in glioblastoma
ZHAO Zhenkai
LIANG Guo
LI Feng
NONG Weixia
ZHANG Qingmei
LUO Bin
XIE Xiaoxun
Abstract：Objective To explore the effect of cancer-testis antigen OY-TES-1 expression on the polarization of M2 macrophages in glioblastoma(GBM).Methods Human GBM cell U251 and human peripheral blood mononuclear cell THP-1 were cultured in vitro to construct a stable transfection cell line U251 that stably down-regulated OY-TES-1(U251-SH-OY-TES-1 group),and U251 cells transfected with unrelated sequences were used as the controls(U251-SH-NC group).Forty-eight hours later,the supernatant of the two groups was co-cultured with THP-1 cells.THP-1 cells to which the supernatant of the U251-SH-NC group was added were taken as the SH-NC group,and THP-1 cells to which the supernatant of the U251-SH-OY-TES-1 group was added were taken as the SH-OY-TES-1 group.The relative expres-sion levels of OY-TES-1 mRNA and protein in the two groups were detected by using real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and Western blotting,respectively,and the transfection efficiency was determined.The expression levels of human interleukin-10(IL-10)and human transforming growth factor-β(TGF-β)in the culture solution were detected by enzyme-linked immunosorbent assay(ELISA).The expression level of CD206 was detected by flow cytometry.Results The detection results of RT-qPCR and Western blotting method showed that the relative expressions of OY-TES-1 mRNA and protein in the U251-SH-OY-TES-1 group were lower than those in the U251-SH-NC group,and the differences were statistically significant(P＜0.05).The results of ELISA showed that the levels of TGF-β and IL-10 in the culture solution of the two groups increased with the increase of culture time.On day 0 and day 2,the levels of TGF-β in the culture solution of the SH-OY-TES-1 group were significantly lower than those of the SH-NC group(P＜0.05),and there were no significant differences in the levels of IL-10 on day 0 and day 2 between the two groups(P＞0.05).On day 4,the levels of TGF-β and IL-10 in the culture solution of the SH-OY-TES-1 group were significantly lower than those of the SH-NC group(P＜0.05).The results of flow cytometry showed that compared with that in the SH-NC group,the level of CD206 in the SH-OY-TES-1 group decreased,and the difference was statisti-cally significant(P＜0.05).Conclusion The high expression of OY-TES-1 in GBM can promote the polarization of M2 macrophages.
Keywords：Glioblastoma(GBM)OY-TES-1M2 macrophagePolarization
Publication Date：2024-08-28
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 875-880 )