乳腺癌组织中整体m6A修饰水平与YTHDF3基因的表达  

Global m6A modification level and YTHDF3 expression in breast cancer tissue

作　　者：孙婷婷 方文 熊伟 王伟 邵春燕 SUN Tingting;FANG Wen;XIONG Wei;WANG Wei;SHAO Chunyan(Department of Clinical Biochemistry,School of Clinical Laboratory Science,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Medical Laboratory,the Affiliated Cancer Hospital of Guizhou Medical University,Guiyang 550001,Guizhou,China)

机构地区：[1]贵州医科大学医学检验学院临床生物化学教研室,贵州贵阳550004 [2]贵州医科大学附属肿瘤医院医学检验科,贵州贵阳550001

出　　处：《贵州医科大学学报》2024年第10期1447-1454,共8页Journal of Guizhou Medical University

基　　金：贵州省科技计划项目(黔科合基础ZK〔2022〕一般419)。

摘　　要：目的探讨乳腺癌组织中mRNA整体N6-甲基腺苷(m6A)修饰水平和甲基化阅读蛋白关键基因YTHDF3基因在乳腺癌组织和癌旁组织中的表达。方法通过生物信息学对YTHDF3基因在乳腺癌组织与癌旁组织中的表达差异及乳腺癌患者生存率进行分析;收集乳腺外科手术切除的25例乳腺癌组织和20例癌旁组织(作为对照),利用液相色谱串联质谱(LC-MS/MS)方法分析乳腺癌组织与癌旁组织组中mRNA的整体甲基化水平,通过实时荧光定量PCR(qPCR)和Western blot实验检测YTHDF3基因在乳腺癌组织和癌旁组织中的相对表达量。结果生物信息数据库结果显示,YTHDF3基因在乳腺癌组织中的表达明显高于癌旁组织(P<0.001),与癌旁组织比较,YTHDF3基因只有在和81~100岁年龄段表达差异具有统计学意义(P<0.05),乳腺癌旁组织中YTHDF3基因的表达量明显低于Luminal分型的乳腺癌(P<0.01),利用受试者工作特征(ROC)曲线验证乳腺癌5年生存率和10年生存率曲线下面积(AUC)>0.56、0.60,高表达YTHDF3基因的乳腺癌患者比低表达的患者存活率低((P<0.05);LC-MS/MS结果显示,乳腺癌分子分型中Luminal型的mRNA的整体甲基化水平高于癌旁组织、差异有统计学意义(P<0.05),HER-2过表达型(HER-2 positive)与癌旁组织的整体甲基化水平差异无统计学意义(P>0.05);qPCR检测结果显示,YTHDF3基因的表达量在乳腺癌分子Luminal亚型、HER-2 positive亚型中均高于癌旁组织(P<0.05);Western blot结果显示,乳腺癌中YTHDF3蛋白水平高于癌旁组织(P<0.05)。结论乳腺癌组织中mRNA的甲基化水平高于癌旁组织,并且甲基化阅读蛋白关键基因YTHDF3基因在乳腺癌组织中表达上调。Objective To investigate global mRNA N-6methyladenosine(m6A)modification level in breast cancer tissues and the expression of YTHDF3,a key m6A reader,in breast cancer tissues and adjacent tissues.Methods Bioinformatics was performed to analyze the difference of YTHDF3 expression between breast cancer tissues and adjacent tissues and the survival rate of patients with breast cancer.Clinical samples were collected from breast surgery,including 25 cases of breast cancer tissues and 20 cases of adjacent tissues(as control).Liquid chromatography tandem mass spectrometry(LC-MS/MS)was applied to analyze the global mRNA methylation level in breast cancer tissues and adjacent tissues.Real-time quantitative PCR(qPCR)and Western blot were used to detect the relative expression of YTHDF3 gene in breast cancer tissues and adjacent tissues.Results Bioinformatics analysis showed that YTHDF3 expression in breast cancer tissues was significantly higher than that in adjacent tissues(P<0.001).YTHDF3 expression in breast cancer tissues was only significantly different from that in adjacent tissues in the patients aged 81-100 years(P<0.05).YTHDF3 expression in adjacent tissues was significantly lower than that in luminal type of breast cancer tissues(P<0.01).Receiver Operating Characteristic(ROC)curve was used to verify that the 5-and 10-year survival rates.Areas under ROC(AUC)of 5-and 10-year survival rates were greater than 0.56 and 0.60,respectively.The survival rate of breast cancer patients with highly expressed YTHDF3 was lower than that of breast cancer patients with lowly expressed YTHDF3(P<0.05).LC-MS/MS results showed that the global mRNA methylation level in luminal type of breast cancer was higher than that in adjacent tissues(P<0.05).There was no statistically significant difference in the global methylation level between HER-2 positive type and adjacent tissues(P>0.05).qPCR results showed that YTHDF3 expressions in luminal and HER-2 positive types in breast cancer were higher than that in adjacent tissues(P<0.05).Wester

关 键 词：乳腺癌 N6-甲基腺苷修饰 液相色谱串联质谱 YTHDF3基因 实时荧光定量PCR 

分 类 号：R735.2[医药卫生—肿瘤]