儿童伯基特淋巴瘤中差异表达基因的鉴定及临床应用

Identification of differentially expressed target genes in pediatric Burkitt lymphoma and its clinical application

目的:分析儿童伯基特淋巴瘤(Burkitt lymphoma,BL)组织与正常淋巴结组织中磷脂酰肌醇3-激酶(phosphatidylinositide3-kinase,PI3K)信号通路、核因子κB(nuclear factor kappa B,NF-κB)信号通路及RAS/RAF信号通路等相关基因在mRNA及蛋白水平上的表达差异,探讨这些基因及相关信号通路与BL发生、发展间的关系,鉴定可用于BL临床诊断、治疗的特异性分子靶标。方法:收集18例儿童BL的组织切片和20例无癌变儿童淋巴结组织(对照)切片,用实时荧光定量反转录聚合酶链反应(reverse transcription polymerase chain reaction,RT-PCR)分析上述信号通路相关基因mRNA水平的表达情况,并用免疫组组化学技术进行蛋白水平表达的分析验证。结果:与对照相比,MYC、TCF3、ID3、CCDN3、CDK6、CDK4、NRAS、RAF1等基因及相应的蛋白在肿瘤组织中高表达,而一些NF-κB信号转导通路目的基因(如BCL2、CD44、CCND2、C-FLIP、A20)表达则受到抑制(P<0.05)。c-MYC基因易位伴BCL2表达(BCL2+)的儿童BL患者其血清乳酸脱氢酶(lactate dehydrogenase,LDH)水平升高(P<0.05)。结论:MYC、TCF3、CCND3、CDK6、NRAS和RAF1等基因在儿童BL中高表达,可作为临床BL诊断和治疗监控的分子靶点;c-MYC基因易位伴BCL2+在儿童BL中与高LDH水平相关,其提示患儿具有较晚的肿瘤分期、较大的肿瘤负荷及不良的预后,可作为临床辅助诊断BL的重要分子指标。

Objective:To identify the differentially expressed genes of phosphatidylinositide3-kinase(PI3 K),nuclear factor kappa-B(NF-κB)and RAS/RAF signal pathways at mRNA and protein levels in pediatric Burkitt lymphoma(BL)for exploring the correlation of these genes with the development and progression of BL and determining the specific target genes for diagnosis and treatment monitoring.Methods:Tumor tissues from 18 cases of pediatric BL and tissues of normal lymphnodes of 20 pediatric cases were collected.The expression levels of m RNA of related target genes in the pathways were analyzed by real-time fluorescence quantitative reverse transcription polymerase chain reaction and the protein levels were determined by immunohistochemistry.Results:The expressions of MYC,TCF3,ID3,CCDN3,CDK6,CDK4,NRAS,RAF1 at mRNA and protein levels were up-expressed significantly in BL and some target genes and its protein of NF-κB pathway such as BCL2,CD44,C-FLIP,CCND2 and A20 were suppressed in BL when compared with normal tissues(P<0.05).Cases carrying c-MYC translocation and BCL2 expression(BCL2+)had higher serum LDH levels(P<0.05).Conclusions:Expressions of MYC,TCF3,CCND3,CDK6,NRAS and RAF1 genes are high,which might be used as molecular markers for the diagnosis and treatment monitoring of pediatric BL.c-MYC gene translocation accompanied with BCL2+might be an important molecular indicator in BL,which is correlated with high serum LDH level,indicating advanced tumor staging,high tumor burden,and poor prognosis.