NF-κB信号通路介导CRY1基因甲基化促进急性髓系白血病预后的机制研究  

Mechanism of NF-KB Signaling Pathway Mediated CRY1 Gene Methylation to Promote Prognosis of Acute Myeloid Leukemia

作　　者：蒋明 王煜煌 陈佳怡 刘蓬勃 葛欣晔 陈蕊 李晨欣 吴明彩[2] J LANG Ming;WANG Yuhuang;CHEN Jiayi;LIU Pengbo;GE Xinye;CHEN Rui;LI Chenucin;WU Mingcai(Wuhu Second Sanatorium for Retired Cadres,Anhui Prouincial Military Command,Wuhu Anhui 241002.China)

机构地区：[1]安徽省军区芜湖市第二离职干部休养所,安徽芜湖241002 [2]皖南医学院生物化学与分子生物学教研室

出　　处：《联勤军事医学》2023年第7期559-563,584,共6页Military Medicine of Joint Logistics

基　　金：国家级大学生创新创业训练计划项目(202210368035、202210368054);安徽省高等学校优秀青年人才支持项目(gxyq2020025);安徽省大学生创新创业训练计划项目(S202110368127)。

摘　　要：目的 检测急性髓系白血病(acute myeloid leukemia, AML)中隐色素1(cryptochrome 1,CRY1)甲基化状态,探讨CRY1甲基化与AML预后的相关性,进一步探讨其促进AML预后的相关机制。方法 培养THP-1细胞,收集58例AML患者(AML组)及23例正常健康人(正常组)骨髓标本,进行基因组DNA及RNA提取。甲基化特异性聚合酶链反应(methylation specific polymerase chain reaction, MSP)方法对CRY1基因进行甲基化检测。逆转录实时荧光定量聚合酶链反应(reverse transcription quantitative real-time polymerase chain reaction, RT-qPCR)和Western blot分别对CRY1、核因子κB p65(nuclear factor kappa-B p65,NF-κB p65)、磷酸化核因子κB p65(phospho-nuclear factor kappa-B p65,p-NF-κB p65)及核因子κB抑制因子α(inhibitor of NF-κB,IκBα)基因和蛋白的表达进行检测。结果 AML组患者的CRY1启动子甲基化率高于正常组(P<0.05)。异常核型AML患者CRY1基因甲基化率和CRY1 mRNA表达分别低于和高于正常核型AML患者。CRY1的甲基化阳性率在不同年龄、性别和FAB分型方面差异无统计学意义(P>0.05)。CRY1甲基化组CRY1、NF-κB p65、p-NF-κB p65及IκBα的mRNA和蛋白相对表达量较CRY1未甲基化组均明显下降(P均<0.05)。结论 AML患者CRY1启动子甲基化下调CRY1 mRNA表达,通过抑制NF-κB的抗凋亡作用,影响细胞增殖与凋亡,促进AML的预后。Objective To detect the methylation of cryptochrome 1(CRY1)in acute myeloid leukemia(AML),to investigate the correlation between CRY1 methylation and prognosis in AML,and to further explore the mechanism of its promotion of prognosis in AML.Methods THP-1 cells were cultured,bone marrow samples from 58 AML patients(AML group)and 23 normal healthy people(normal group)were collected for extraction of genomic DNA and RNA.Methylation specific polymerase chain reaction(MSP)was used to detect CRY1 gene methylation.CRY1,nuclear factor kappa-B p65(NF-kB p65),phospho-nuclear factor kappa-B p65(p-NF-kB p65)and inhibitor of NF-B(IkBα)gene and protein expression were detected by reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR)and Western blot respectively.Results Methylation rate of CRY1 promoter in AML group was higher than that in normal group(P<0.05).The CRY1 gene methylation rate and CRY1 mRNA expression in abnormal karyotype AML patients were lower and higher than those in normal karyotype AML patients respectively.There was no significant difference in the methylation positive rate of CRY1 among different age,sex and FAB types(P>0.05).The mRNA and protein relative expressions of CRY1,NF-sB p65,p-NF-kB p65 and IkBαin CRY1 methylated group significantly decreased compared with those in CRY1 unmethylated group(all P<0.05).Conclusion CRY1 promoter methylation down-regulates CRY1 mRNA expression in AML patients,affects cell proliferation and apoptosis by inhibiting the anti-apoptotic effects of NFB,and promotes the prognosis of AML patients.

关 键 词：NF-ΚB信号通路 隐色素1 甲基化 急性髓系白血病 

分 类 号：R733.71[医药卫生—肿瘤]