摘要
目的:探讨SOCS1基因的甲基化状态与JAK2/STAT信号通路的活性及急性髓系白血病发生发展的关系。方法:采用细胞培养、qPCR、MS-PCR、Western blot、CCK-8检测、流式细胞术及基因转染等技术,分析120例急性髓系白血病患者及白血病细胞株U937和THP-1中SOCS1基因的表达、甲基化状态与疾病发生发展的关系。同时分析SOCS1下游JAK2/STAT信号通路各蛋白的变化及对白血病细胞株生长和凋亡的影响。结果:在SOCS1甲基化组WT1/ABL的阳性率显著高于阴性组,治疗1个疗程完全缓解率明显低于阴性组。在SOCS1基因甲基化率低组中该基因的表达较高,其下游p-JAK2、p-STAT3和p-STAT5表达降低,t-JAK2、t-STAT3和t-STAT5的变化无显著差异。随着去甲基化药物浓度的增加,白血病细胞生长率逐渐降低,细胞凋亡率逐渐增高。结论:SOCS1基因的甲基化导致基因表达沉默,降低其对JAK2/STAT通路信号传导的抑制,最终促进急性髓系白血病细胞的生长和增殖。
Objective:To explore the correlation of SOCS1 gene methylation with the activity of JAK2/STAT signaling pathway,genesis and progress of acute myeloid leukemia.Methods:Several techniques,such as cell culture,qPCR,MS-PCR,Western bolt,CCK-8 assay,flow cytometry and gene transfection were used to analyze the relation of expression and methylation statues of SOCS1 gene with the genesis and progression of AML in 120 AML patients and leukemia cell lines U937 and THP-1,at the same time to analyze the changes of downstream protein expression in JAK2/STAT signaling pathway and their effect on the growth and apoptosis of leukemia cell lines.Results:The positive rate of WT1/ABL in SOCS1 methylated group was significantly higher than that in SOCS1 non-methylated group,and the complete remission rate of one course treatment in SOCS1-methylated group was significantly lower than that in SOCS1 non-methylated group.The expression level of SOCS1 gene in low methylation rate group was higher,and the expression of down-stream proteins p-JAK2,p-STAT3 and p-STAT5 in JAK2/STAT signaling pathway decreased,while the expression of t-JAK2,t-STAT3 and t-STAT5 was not changed statistically significantly.The growth rate of leukemia cells graduated decreased,and the apoptosis rate of leukemia cells graduated increased along with the enhancement of methylation drug concentration.Conclusion:The methylation of SOCS1 gene results in the gene silencing,thereby declines its inhibition on the down-stream proteins in JAK2/STAT signaling pathway,and finally promotes the growth and proliferation of AML cells.
作者
张晓慧
罗建民
杨琳
刘小军
ZHANG Xiao-Hui;LUO Jian-Min;YANG Lin;LIU Xiao-Jun(Department of Hematology,Handan Central Hospital of Hebei Province,Handan 056001,Hebei Province,China;Department of Hematology,The Second Hospital of Hebei Medical University,Shijiazhuang 050000,Hebei Province,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2020年第5期1496-1503,共8页
Journal of Experimental Hematology
基金
河北省卫生健康委员会科研基金(20190195)。
