髓系白血病细胞中细胞因子信号转导抑制因子1基因启动子去甲基化对细胞增殖、凋亡的影响  

Effects of suppressor of cytokine signaling 1 gene promoter demethylation on cell proliferation and apoptosis in myeloid leukemia cells

作　　者：陈莉[1] 李青山[2] 张学强[3] 任利彬[4] 索晓慧 张丛丛[1] CHEN Li;LI Qingshan;ZHANG Xueqiang;REN Libin;SUO Xiaohui;ZHANG Congcong(the First Department of Hematology,Handan Central Hospital in Hebei Province,Handan,Hebei,056001;Department of Orthopedics,Handan Central Hospital in Hebei Province,Handan,Hebei,056001;Department of Oral and Maxillofacial Surgery,Handan Central Hospital in Hebei Province,Handan,Hebei,056001;Department of General Surgery,Handan Central Hospital in Hebei Province,Handan,Hebei,056001)

机构地区：[1]河北省邯郸市中心医院血液内一科,河北邯郸056001 [2]河北省邯郸市中心医院骨科,河北邯郸056001 [3]河北省邯郸市中心医院口腔颌面外科,河北邯郸056001 [4]河北省邯郸市中心医院普外科,河北邯郸056001

出　　处：《实用临床医药杂志》2022年第8期60-65,共6页Journal of Clinical Medicine in Practice

基　　金：河北省邯郸市科学技术研究与发展计划项目(19422083009-24)。

摘　　要：目的探讨髓系白血病(ML)细胞中细胞因子信号转导抑制因子1(SOCS-1)基因启动子去甲基化对细胞增殖、凋亡的影响。方法收集78例ML患者的骨髓标本(实验组)以及人ML细胞系U937、HL-60、K562为研究对象,另外收集50例健康捐献者的骨髓标本作为对照组。甲基化特异性聚合酶链反应(MS-PCR)检测ML患者骨髓标本中SOCS-1基因启动子甲基化状态,实时荧光定量聚合酶链反应(qRT-PCR)检测ML患者骨髓标本以及人ML细胞系中SOCS-1 mRNA表达;Western blot检测ML患者骨髓标本以及人ML细胞系中SOCS-1蛋白表达。用0、0.8、1.6、3.2μmol/L地西他滨(DCA)分别处理U937细胞,并分别命名为空白组、DCA-L组(DCA低剂量)、DCA-M组(DCA中剂量)、DCA-H组(DCA高剂量)。MS-PCR检测各组细胞中SOCS-1基因启动子甲基化状态;qRT-PCR检测各组细胞中SOCS-1 mRNA表达;Western blot检测各组细胞中SOCS-1蛋白表达;CCK-8法检测各组细胞增殖情况;流式细胞术检测各组细胞凋亡情况。结果SOCS-1在ML患者中呈高甲基化状态,SOCS-1 mRNA及SOCS-1蛋白在ML患者中的表达低于对照组,差异有统计学意义(P<0.05);人ML细胞系U937、HL-60、K562中SOCS-1 mRNA及SOCS-1蛋白表达低于人骨髓基质细胞HS-5,差异有统计学意义(P<0.05),且U937细胞中SOCS-1 mRNA及SOCS-1蛋白表达水平最低,因此,以U937细胞为研究对象进行后续实验。与空白组比较,DCA-L组、DCA-M组、DCA-H组中SOCS-1甲基化水平、450 nm处光密度值(OD_(450 nm))(24、48、72 h)降低,SOCS-1 mRNA及SOCS-1蛋白表达水平、细胞凋亡率升高(P<0.05),且随着DCA浓度的升高,上述指标的相应趋势越明显。结论SOCS-1去甲基化可抑制ML细胞增殖、促进细胞凋亡。Objective To investigate the effects of demethylation of the promoter of suppressor of cytokine signaling 1(SOCS-1)in myeloid leukemia(ML)cells on cell proliferation and apoptosis.Methods The bone marrow specimens of 78 ML patients(experimental group)and the human ML cell lines U937,HL-60 and K562 were collected as research objects,and bone marrow specimens from 50 healthy donors were collected as control group.Methylation-specific polymerase chain reaction(MS-PCR)was used to detect the methylation status of SOCS-1 gene promoter in bone marrow specimens of ML patients,quantitative real-time polymerase chain reaction(qRT-PCR)was used to detect the expression of SOCS-1 mRNA in bone marrow specimens of ML patients and human ML cell lines;Western blot was used to detect the expression of SOCS-1 protein in bone marrow specimens of ML patients and human ML cell lines.U937 cells were treated with 0,0.8,1.6 and 3.2μmol/L Decitabine(DCA),and were selected as blank group,DCA-L group(DCA low-dose),DCA-M group(DCA medium-dose)and DCA-H group(DCA high-dose).MS-PCR was used to detect the SOCS-1 gene promoter methylation status in each group of cells;qRT-PCR was used to detect the SOCS-1 mRNA expression in cells of each group;western blot was used to detect the SOCS-1 protein expression in each group of cells;CCK-8 method was used to detect the cell proliferation in each group;flow cytometry was used to detect the cell apoptosis in each group.Results SOCS-1 was hypermethylated in ML patients,and the expressions of SOCS-1 mRNA and SOCS-1 protein in ML patients were significantly lower than that in the control group(P<0.05);the expressions of SOCS-1 mRNA and SOCS-1 protein in human ML cell lines U937,HL-60,and K562 was significantly lower than that in human bone marrow stromal cells HS-5(P<0.05),and the expression levels of SOCS-1 mRNA and SOCS-1 protein in U937 cells were the lowest,therefore,the U937 cells were used as the research objects for follow-up experiments.Compared with the blank group,the SOCS-1 methylation levels an

关 键 词：髓系白血病 细胞因子信号转导抑制因子1 甲基化 细胞增殖 细胞凋亡 

分 类 号：R733.7[医药卫生—肿瘤] Q81[医药卫生—临床医学]