【摘要】 目的探讨肝癌缺失基因1(deleted in liver cancer-1,DLC-1)启动子在视网膜母细胞瘤(retinoblastoma,RB)中的甲基化情况及与临床参数的关系。方法收集37例RB石蜡切片,4例正常眼部组织标本和2株RB细胞系(WERI-Rb1及Y79)。用甲基化敏感性高分辨率熔解(MS-HRM)法定量检测DLC-1甲基化情况,用亚硫酸氢盐测序对其进行验证,并用RT-PCR检测WERI-Rb1和Y79细胞系中DLC-1 mRNA的表达。结果 35.1%的RB中存在DLC-1启动子甲基化,而在正常眼部组织中未发现甲基化;WERI-Rb1和Y79检测出DLC-1 mRNA表达。DLC-1启动子甲基化与肿瘤分化、伴随其他肿瘤和家族史均无显著关系,但单侧RB患者中的DLC-1甲基化率明显高于双侧患者(P<0.05)。结论 RB患者DLC-1启动子存在一定程度的甲基化,DLC-1甲基化在RB的发生、发展中具有一定的作用。更多还原

【Abstract】 Objective To investigate the methylation status of DLC-1 in retinoblastoma(RB) tissue and its correlation with clinical parameters.Methods Thirty-seven paraffin-embedded RB tissue samples,4 normal eye tissue samples and 2 retinoblastoma cell line(Y79 and WERI-Rb1) samples were collected,and their methylation status of DLC-1 were quantitatively detected with the methylation-sensitive high resolution melting(MS-HRM) method.Then,the bisulfite genomic sequencing was used to validate the methylation status of DLC-1 gene.In the meanwhile,the expression of DLC-1 mRNA in Y79 and WERI-Rb1 cell lines were determined by RT-PCR.Results Aberrant DLC-1 methylation was detected in 13(35.1%) of 37 RB samples,but was not found in normal eye tissues.DLC-1 mRNA was detected in Y79 and WERI-Rb1 cell lines.The DLC-1 methylation was not correlated with tumor differentiation,other accompanied cancers and family history.However,the methylation rate of DLC-1 promoter in unilateral RB patients was significantly higher than that in bilateral patients(P<0.05).Conclusion Aberrant methylation of DLC-1 promoter exists in RB patients,and may play a role in the pathogenesis of RB.更多还原