EZH2抑制剂GSK343对脑胶质瘤细胞EZH2/STAT4/IFN-γ通路及γδT细胞杀伤作用的影响  

Influences of GSK343 on EZH2/STAT4/IFN-γPathway and the Killing Effect ofγδT Cells in Glioma Cells

作　　者：夏丽琼[1] 杨明华[1] 谢燕[1] XIA Liqiong;YANG Minghua;XIE Yan(Mianyang Central Hospital, Sichuan Mianyang 621000, China)

机构地区：[1]四川省绵阳市中心医院呼吸与危重症医学科,四川绵阳621000

出　　处：《河北医学》2022年第6期904-909,共6页Hebei Medicine

基　　金：中国宋庆龄基金会呼吸疾病临床研究工艺基金慢肺阻专项定向委托研究课题任务,(编号:2018MZFS-046)。

摘　　要：目的:探讨EZH2抑制剂GSK343对脑胶质瘤细胞Zeste基因增强子人类同源物2(EZH2)/信号转导与转录激活因子4(STAT4)/干扰素-γ(IFN-γ)通路及γδT细胞杀伤作用的影响。方法:体外培养人脑胶质瘤细胞系U373,设置对照组(细胞正常培养,不做药物处理)和GSK343组(GSK343处理细胞)。CCK-8法检测各组U373细胞增殖情况及γδT细胞对U373细胞的杀伤作用;流式细胞仪检测各组U373细胞凋亡情况;显微镜观察细胞形态;蛋白印迹(WB)法检测各组U373细胞中EZH2、p-STAT4、IFN-γ蛋白表达情况。结果:随着GSK343的处理及处理浓度的升高,U373细胞增殖抑制率逐渐升高,U373细胞的GSK343半数抑制浓度(IC50)处于20-50μmoL/L范围内,故选择50μmoL/L作为GSK343组的最佳处理浓度进行后续实验;与对照组相比,GSK343组EZH2显著降低(P<0.05),细胞凋亡率、p-STAT4、IFN-γ蛋白表达水平、各个时间点γδT细胞对U373细胞的杀伤活性显著升高(P<0.05),γδT细胞杀伤的形态不规则、胞质肿胀的U373细胞增多。结论:GSK343可抑制EZH2蛋白表达,促进STAT4蛋白磷酸化、IFN-γ蛋白表达,从而增强γδT细胞对U373细胞的杀伤作用,抑制U373细胞增殖并诱导其凋亡。Objective:To investigate the influences of GSK343 on the enhancer of Zeste homolog 2(EZH2)/signal transducer and activator of transcription 4(STAT4)/interferon-γ(IFN-γ)pathway and the killing effect ofγδT cells.Methods:Human glioma cell line U373 was cultured in vitro,and the control group(normal cell cultured,no drug treatment)and GSK343 group(GSK343 treatment)were set up.The proliferation of U373 cells and the killing effect ofγδT cells on U373 cells were detected by CCK-8 method;the apoptosis of U373 cells was detected by flow cytometry;the cell morphology was observed by microscopy;and the expressions of EZH2,p-STAT4 and IFN-γproteins in U373 cells were detected by Western blot.Results:With the increase of GSK343 concentration,the inhibition rate of U373 cell proliferation increased gradually,and the GSK343 half inhibition concentration(IC50)of U373 cells was in the range of 20-50μmol/L.Therefore,50μmol/L were selected as the best treatment concentration in the GSK343 group for subsequent experiments.Compared with the control group,the expression levels of EZH2 protein in GSK343 group decreased significantly(P<0.05),and the apoptosis rate,p-STAT4,IFN-γprotein expression level and the killing activity ofγδT cells to U373 cells in each group were significantly increased(P<0.05),irregular morphology and cytoplasmic swelling U373 cells killing ofγδT cells increased.Conclusion:GSK343 can inhibit the expressions of EZH2 proteins,promote the expression of STAT4 phosphorylation and IFN-γprotein,thus enhancing the killing effect ofγδT cells on U373 cells,inhibit the proliferation and induce apoptosis of U373 cells.

关 键 词：GSK343 脑胶质瘤 Zeste基因增强子人类同源物2 信号转导与转录激活因子4 干扰素-Γ ΓΔT细胞 

分 类 号：R73[医药卫生—肿瘤]