CAJ | 학술논문

目的探讨肾母细胞瘤患儿血液中SIX2基因的转录表达及其启动子甲基化状态以及其与临床病理学特征的关系.方法收集自2008年10月至2012年1月入住郑州大学第一附属医院小儿外科的45例肾母细胞瘤患儿作为病例组(男29例、女16例),以健康体检或就诊的性别、年龄匹配的排除肿瘤等恶性疾病患儿15例作为对照组.采集所有研究对象外周血,运用实时荧光定量PCR(qRT-PCR)和甲基化特异性PCR (MSP)法检测SIX2基因转录表达水平及其启动子甲基化状态,t或x2检验比较组间差异,并分析病例组中两者与临床病理特征的关系及SIX2基因的甲基化对其转录表达的影响.结果病例组中SIX2 mRNA的相对表达量(RQ)高于对照组(1.93±1.10比0.57±0.39,t=5.354,P=0.000)；病例组SIX2基因甲基化阳性者有8例,对照组12例,病例组中甲基化阳性比率更低(x2 =19.600,P=0.000).病例组的RQ值与肿瘤大小、临床分期、病理分型、淋巴转移、治疗转归均有关(均P ＜0.05).病例组和对照组中甲基化组的RQ值均低于非甲基化组(1.35±0.44比1.95±1.15,0.43±0.29比1.13±0.20,t=2.459、3.896,P=0.020、0.002)；甲基化组中病例组的RQ值高于对照组(t=5.624,P=0.000)；非甲基化组中两者之间的RQ值比较差异无统计学意义(t=1.222,P=0.229).结论肾母细胞瘤外周血中SIX2基因的异常甲基化与转录表达有关,是基因表达调节方式之一,并与肾母细胞瘤的发生发展有关；SIX2基因在发生甲基化的肾母细胞瘤中有可能充当癌基因的角色,转录水平的过表达与其甲基化状态有负相关倾向.
목적 탐토신모세포류환인혈액중SIX2기인적전록표체급기계동자갑기화상태이급기여림상병이학특정적관계.방법 수집자2008년10월지2012년1월입주정주대학제일부속의원소인외과적45례신모세포류환인작위병례조(남29례、녀16례),이건강체검혹취진적성별、년령필배적배제종류등악성질병환인15례작위대조조.채집소유연구대상외주혈,운용실시형광정량PCR(qRT-PCR)화갑기화특이성PCR (MSP)법검측SIX2기인전록표체수평급기계동자갑기화상태,t혹x2검험비교조간차이,병분석병례조중량자여림상병리특정적관계급SIX2기인적갑기화대기전록표체적영향.결과 병례조중SIX2 mRNA적상대표체량(RQ)고우대조조(1.93±1.10비0.57±0.39,t=5.354,P=0.000)；병례조SIX2기인갑기화양성자유8례,대조조12례,병례조중갑기화양성비솔경저(x2 =19.600,P=0.000).병례조적RQ치여종류대소、림상분기、병리분형、림파전이、치료전귀균유관(균P ＜0.05).병례조화대조조중갑기화조적RQ치균저우비갑기화조(1.35±0.44비1.95±1.15,0.43±0.29비1.13±0.20,t=2.459、3.896,P=0.020、0.002)；갑기화조중병례조적RQ치고우대조조(t=5.624,P=0.000)；비갑기화조중량자지간적RQ치비교차이무통계학의의(t=1.222,P=0.229).결론 신모세포류외주혈중SIX2기인적이상갑기화여전록표체유관,시기인표체조절방식지일,병여신모세포류적발생발전유관；SIX2기인재발생갑기화적신모세포류중유가능충당암기인적각색,전록수평적과표체여기갑기화상태유부상관경향.
Objective To explore the transcriptional expression and promoter methylalion status of SIX2 gene in peripheral blood of pediatric children with nephroblastoma and discuss their clinicopathological correlations.Methods Approved by the hospital ethics committee,peripheral blood samples were collected from 45 children with Wilms' tumor(case group) at the Department of Pediatric Surgery,First Affiliated Hospital,Zhengzhou University from October 2008 to January 2012.And another 15 pediatric cases genderand-age matched,were selected as the control group (excluding cancer and other malignant diseases).The real-time quantitative (qRT)-PCR and methylation-specific PCR (MSP) were used to detect the mRNA expression level and methylation status of SIX2 gene.t or x2 test were used.Then analyzed their clinicopathological correlations in the case group and how SIX2 gene methylation affected its transcription.Results Relative quantity(RQ) of SIX2 mRNA in the case group was higher than that of the control group (1.93 ± 1.10 vs 0.57 ± 0.39,t =5.354,P =0.000).There were 8 SIX2 gene methylation-positive cases in the case group versus 12 cases in the control group.And the methylation positive ratio was extremely lower in the case group (x2 =19.600,P =0.000).RQ values in the case group was associated with tumor size,clinical stage,pathological type,lymph node metastasis,treatment and outcome (all P ＜ 0.05).RQ values in the methylated group was lower than that of the unmethylated group both in case and control group(1.35 ±0.44vs 1.95 ±1.15,0.43 ±0.29 vs 1.13 ±0.20,t=2.459 and 3.896,P=0.020 and 0.002).RQ values of case group was higher than that of the control group in methylated group(t =5.624,P =0.000).No statistical significance existed in RQ values between the case and control groups of unmethylated group (t =1.222,P =0.229).Conclusions A close correlation between SIX2 low methylation and high mRNA expression in blood suggests that aberrant promoter methylation is possibly one of gene expression regulations,and may be correlated with the occurrence and development of Wilms' tumor.And SIX2 gene in methylated Wilms' tumor may play the role of oncogenes.A negative correlation exists between the overexpression in transcriptional level and its methylation status.