儿童视网膜母细胞瘤发生发展密切相关的可疑新基因位点研究(英文)

New suspicious gene loci correlated with the generation and development of children's retinoblastoma

背景:近年来研究表明视网膜母细胞瘤(retinoblastoma,RB)的发生和病变的进展除了与已知的Rb1基因有相关性外,可能还有其他抑癌基因的参与。目的:探讨其他可能参与RB发生发展的基因存在的位点,并试图寻找和确定具有监测及预后价值的杂合性缺失(LOH)检测指标。设计:以RB患者为研究对象的病例分析。单位:一所军医大学医院的基因诊断治疗中心。对象:本研究在第三军医大学西南医院基因诊断治疗中心完成,研究对象为1998-05/2001-10在本校三所附属医院门诊就诊的RB患者16例。纳入标准:符合RB诊断标准的年龄小于3岁的患儿;排除标准:有家族遗传史者。其中男10例,女6例;累及双眼者12例,累及单眼者4例。方法:在患者的RB肿瘤及外周血标本中运用荧光聚合酶链反应(PCR)分别扩增13号染色体上14个微卫星DNA标记,分析测定各位点LOH发生率;同时通过家系分析确定位点缺失的遗传学来源。主要观察指标:患者13号染色体上LOH发生频率。结果:16例RB患者中,12例在13号染色体上1个或1个以上位点发生LOH。其中3个位点D13S265、D13S263和D13S153(位于Rb1基因内)LOH发生机率最高。12个LOH阳性标本中有10个位点的缺失被确定发生在父系来源的染色体中。LOH阳性及阴性组RB的确诊时间分别为504和1086d,两组相比差异具有显著性意义(t=2.

BACKGROUND:Recent researches have indicated that the generation and development of retinoblastoma(RB) might also be related with other anti oncogenes except the known Rb1 gene.OBJECTIVE:To explore the loci of other genes which possibly participated in RB generation and development and try to find and confirm the indicators for the loss of heterozygosity(LOH) with merits in surveillance and prognosis.DESIGN:A case analysis by employing RB patients as subjectsSETTING:A center of gene diagnosis and therapy of a military medical university affiliated hospitalPARTICIPANTS:The study was conducted in the center of gene diagnosis and therapy of Xinan Hospital affiliated to Third Military Medical University of Chinese PLA.Sixteen RB cases including 10 males and 6 females were the patients of the outpatient department of three affiliated hospitals of the Third Military Medical University of Chinese PLA from May 1998 to October 2001.Inclusion criteria:in accordance with RB diagnostic criteria and younger than 3 years old; Exclusion criteria:family heredity history.Two eyes were involved in 12 cases and one eye was involved in 4 cases.METHODS:Fourteen micro satellite DNA labels on the 13th chromosome in tumor or peripheral blood samples were separately amplified by polymerase chain reaction(PCR) to analyze the incidence of LOH of each locus.Simultaneously,the genetic source of loci loss was confirmed by genealogical analysis.MAIN OUTCOME MEASURES:Frequency of LOH incidence on the 13th chromosome.RESULTS:In 16 RB patients,LOH occurred in one or more than one locus on the 13th chromosome of 12 cases.Thereinto,the probability of LOH occurrence on three loci including D13S265,D13S263 and D13S153(in Rb1 gene) was the highest.Ten loci of LOH in 12 LOH positive samples were confirmed from agnate chromosomes.The RB confirmation of LOH positive group or LOH negative group needed 504 days or 1086 days, which was significantly different(t=2.357,P< 0.05).CONCLUSION:RB confirmation was earlier in LOH positive patients than LOH negative patients.Except the confirmed Rb1 gene,LOH on two loci including D13S263(13q14.1- 14.2) and D13S265 (13q31- 32) also might have certain suggestive effect on early intervention and functional surveillance of RB patients.