P13K/mTOR信号通路参与调控KG1a细胞NKG2D配体的表达  

The expression of NKG2D ligand of KG1a cell with the participation and regulation of P13K/mTOR signaling pathway

作　　者：何川疆 张素杰 刘雯[1] 李泽泳[2] 周慧芳[1] 胡亮杉[2] 张丽萍[1] 许爱敏[1] HE Chuan-jiang;ZHANG Su-jie;LIU Wen(Department of Laboratory,The First People's Hospital of Kashgar District,Kashgar 844000,China;不详)

机构地区：[1]喀什地区第一人民医院检验科,新疆喀什844000 [2]广东省第二人民医院检验科,广东广州510317

出　　处：《中国医学装备》2021年第5期183-187,共5页China Medical Equipment

基　　金：新疆维吾尔自治区自然科学基金(2017D01C006)“阻断PI3K/mTOR通路逆转白血病干细胞免疫抵抗的机制研究”。

摘　　要：目的:自然杀伤细胞活化性受体(NKG2D)配体的低表达是白血病干细胞(LSC)逃避免疫杀伤的关键因素,磷脂酰肌醇3-激酶/雷帕霉素靶蛋白(PI3K/mTOR)信号通路在LSC中高度活化,探讨PI3K/mTOR信号通路是否参与调控LSC中NKG2D配体的表达。方法:采用PI3K/mTOR信号通路抑制剂NVP-BEZ235对人急性骨髓白血病细胞(KG1a细胞)进行干预后,采用细胞技术试剂盒CCK-8检测KG1a细胞增殖,采用流式细胞术检测KG1a细胞周期及凋亡,采用实时荧光定量聚合酶链反应(RT-qPCR)及免疫印迹(Western blot)检测细胞中NKG2D配体MICB、ULBP1、ULBP2和ULBP3的表达。结果:抑制剂NVP-BEZ235显著抑制KG1a细胞的增殖,并将KG1a细胞周期阻滞于细胞周期G0/G1期;NVP-BEZ235抑制剂干预后,KG1a细胞NKG2D配体MICB信使核糖核酸(mRNA)相对表达量显著升高,ULBP1、ULBP2、ULBP3、mRNA及蛋白表达量也显著升高。结论:PI3K/mTOR调控LSC的增殖及周期,且其能够调控NKG2D的表达,该结果为通过PI3K/mTOR抑制靶向治疗耐药的LSC提供了数据支持。Objective:The low expression of the ligand of natural killer cell group 2D(NKG2D)is a key factor that leukemia stem cells(LSC)escapes cytokine induced killer,and PI3K/mTOR signaling pathway is highly activated in LSC.The aim of this study is to investigate whether the PI3K/mTOR signaling pathway is involved in the expression of NKG2D ligand in regulating LSC.Methods:After the PI3K/mTOR signaling pathway inhibitor NVP-BEZ235 was adopted to intervene human acute myelocytic leukemia cell(KGla cell),the cell technique kit(CCK-8)was adopted to detect the proliferation of KGla cell,and flow cytometry(FCM)was adopted to detect the KGla cell cycle and apoptosis,and realtime fluorescence quantification polymerase chain reaction(RT-qPCR)and Western blot(WT)were adopted to detect the expressions of MICB,ULBP1,ULBP2 and ULBP3 of NKG2D ligand in cell.Results:The NVP-BEZ235 inhibitor significantly inhibited the proliferation of KG1a cells,and inhibited the KGla cell cycle at G0/G1 stage.After NVP-BEZ235 inhibitor intervened cell,the relative expression amount of MICB mRNA of NKG2D ligand in KGla cell was significantly increased,and the expression amount of ULBP1,ULBP2,ULBP3,mRNA and protein also were significantly increased.Conclusion:PI3K/mTOR can regulate the LSC proliferation and cycle,and it can regulate the NKG2D expression.This result provides data support for inhibiting drug resistance LSC with targeted therapy by PI3K/mTOR.

关 键 词：PI3K/mTOR信号通路 白血病干细胞 细胞周期 自然杀伤细胞活化性受体(NKG2D)配体 

分 类 号：R733.71[医药卫生—肿瘤]