IGHG1对人急性髓系白血病THP-1细胞增殖、凋亡的影响  

The Effects of IGHG1 on the Proliferation and Apoptosis of Human Acute Myeloid Leukemia THP-1 Cells

作　　者：高鑫[1] 储李婧 陈天平 GAO Xin;CHU Li-Jing;CHEN Tian-Ping(School of Clinical Medicine,Anhui Medical College,Hefei 230031,Anhui Province,China;Department of Hematology and Oncology,Anhui Children′s Hospital,Hefei 230000,Anhui Province,China)

机构地区：[1]安徽医学高等专科学校临床医学院,安徽合肥230031 [2]安徽省儿童医院血液肿瘤科,安徽合肥230000

出　　处：《中国实验血液学杂志》2023年第5期1263-1271,共9页Journal of Experimental Hematology

基　　金：安徽省自然科学基金青年项目(1608085QH218);安徽省教育厅高校优秀人才支持计划项目(gxyq2020236);安徽医学高等专科学校创新团队资助项目(YZ2020TD005)。

摘　　要：目的:探讨免疫球蛋白G1重链恒定区(IGHG1)对急性髓系白血病(AML)细胞系THP-1细胞增殖、凋亡的影响及其可能的作用机制。方法:体外培养人AML THP-1细胞,分为对照(正常培养的THP-1细胞)、pcDNA3.1[转染IGHG1过表达(pcDNA3.1-IGHG1)阴性对照质粒的THP-1细胞]、pcDNA3.1-IGHG1(转染pcDNA3.1-IGHG1的THP-1细胞)、LY364947[转化生长因子-β(TGF-β)/信号转导蛋白(Smad)抑制剂LY36494720μmol/L处理THP-1细胞)]、si-NC[转染IGHG1小干扰RNA(IGHG1-siRNA)阴性对照的THP-1细胞]、si-IGHG1(转染IGHG1-siRNA的THP-1细胞)和si-IGHG1+LY364947(IGHG1-siRNA和LY364947共同处理THP-1细胞)共7组。荧光定量PCR法检测各组THP-1细胞中IGHG1和免疫球蛋白G(IgG)mRNA的表达;CCK-8法检测各组THP-1细胞增殖活力;流式细胞术检测各组THP-1细胞凋亡率和细胞周期变化;蛋白印迹法检测各组THP-1细胞增殖、凋亡及TGF-β/Smad信号通路相关蛋白的表达。结果:与对照组相比,过表达IGHG1后THP-1细胞中IGHG1和IgG mRNA表达、细胞增殖活力、S期的细胞比例、细胞周期蛋白D1(Cyclin D1)、B细胞淋巴瘤-2(Bcl-2)、IgG、TGF-β1、磷酸化Smad3(p-Smad3)/Smad3蛋白表达均显著升高(P<0.05),细胞凋亡率、G_(0)/G_(1)期的细胞比例、p21、Bcl-2相关X蛋白(Bax)、半胱氨酸天冬氨酸蛋白酶-3(Caspase-3)蛋白表达均显著降低(P<0.05)。抑制TGF-β/Smad信号通路或沉默IGHG1后THP-1细胞中IGHG1和IgG mRNA表达、细胞增殖活力、S期的细胞比例、Cyclin D1、Bcl-2、IgG、TGF-β1、p-Smad3/Smad3蛋白表达均显著降低(P<0.05),细胞凋亡率、G_(0)/G_(1)期的细胞比例、p21、Bax、Caspase-3蛋白表达均显著升高(P<0.05);且与沉默IGHG1相比,IGHG1基因沉默和TGF-β/Smad通路抑制共同处理的THP-1细胞中IGHG1和IgG mRNA表达、细胞增殖活力、S期的细胞比例、Cyclin D1、Bcl-2、IgG、TGF-β1、p-Smad3/Smad3蛋白表达均显著降低(P<0.05),细胞凋亡率、G_(0)/G_(1)期的细胞比例、p21、Bax、Caspase-3Objective:To investigate the effects of the immunoglobulin G1 heavy chain constant region(IGHG1)on the proliferation and apoptosis of acute myeloid leukemia(AML)THP-1 cells and its possible mechanism of action.Methods:Human AML THP-1 cells were cultured in vitro and divided into control(normally cultured THP-1 cells),pcDNA3.1[THP-1 cells transfected with IGHG1 overexpression(pcDNA3.1-IGHG1)negative control plasmid],pcDNA3.1-IGHG1(THP-1 cells transfected with plasmid containing pcDNA3.1-IGHG1),LY364947[transforming growth factor-β(TGF-β)/signal transduction protein(Smad)inhibitor LY36494720μmol/L treated THP-1 cells],si-NC[THP-1 cells transfected with IGHG1-small interfering RNA(siRNA)negative control],si-IGHG1(THP-1 cells transfected with IGHG1-siRNA),and si-IGHG1+LY364947(IGHG1-siRNA and LY364947 co-treated THP-1 cells)a total of 7 groups.Fluorescence quantitative PCR was used to detect the expression of IGHG1 and immunoglobulin G(IgG)mRNA of THP-1 cells in each group;CCK-8 was used to detect THP-1 cells proliferation activity;flow cytometry was used to detect THP-1 cells apoptosis and cell cycle in each group;Western blot was used to detect the THP-1 cells proliferation,apoptosis and the expression of TGF-β/Smad signaling pathway related proteins.Results:Compared with the control group,after overexpression of IGHG1,the expression of IGHG1 and IgG mRNA,cell proliferation viability,S phase cell ratio,expressions of Cyclin D1,B cell lymphoma-2(Bcl-2),IgG,TGF-β1,phosphorylated Smad3(p-Smad3)/Smad3 protein in THP-1 cells were significantly increased(P<0.05),the apoptosis rate,G0/G1 phase cell ratio,expression of p21,Bcl-2 related X protein(Bax),Caspase-3 protein were significantly reduced(P<0.05);after inhibiting TGF-β/Smad signaling pathway or silencing IGHG1,the expression of IGHG1 and IgG mRNA,cell proliferation viability,S phase cell ratio,expression of Cyclin D1,Bcl-2,IgG,TGF-β1,p-Smad3/Smad3 protein in THP-1 cells were significantly reduced(P<0.05),the apoptosis rate,G0/G1 phase cell ratio,expressions o

关 键 词：免疫球蛋白G1重链恒定区 急性髓系白血病THP-1细胞 免疫球蛋白G 凋亡 增殖 转化生长因子-β/信号转导蛋白 

分 类 号：R733.71[医药卫生—肿瘤]