TCRP1基因启动子区CpG岛测序体系的建立

Establishment of a method for sequencing CpG islands located at the promoter region of TCRP1 gene

目的建立可用于舌癌耐药蛋白1(TCRP1)基因启动子区CpG岛测序检测的反应体系。方法采用人肺癌A549细胞作为待测样品,以TCRP1启动子区CpG岛为靶片段设计特异扩增、测序引物,建立TCRP1启动子区CpG岛测序检测反应体系,并检测H1299、H460、Huh-7细胞株和2例患者肿瘤组织标本。结果建立了以P2-1[二甲基亚砜(DMSO)终浓度0%,降落聚合酶链反应(PCR)条件]联合P1-2(DMSO终浓度1%,常规PCR反应条件)为优选方案的TCRP1启动子区CpG岛测序检测体系,并成功检测了A549、H1299、H460、Huh-7细胞株和2例患者肿瘤组织标本。结论成功建立可用于TCRP1启动子区CpG岛测序的反应体系。

Objective To establish a method for sequencing the CpG islands located at promoter region of tongue cancer-resistant protein 1（TCRP1）gene.Methods A method for sequencing the CpG islands located at the promoter region of TCRP1 gene was established with specific polymerase chain reaction（PCR）and sequencing primers by using human cell line A549.Then human cell line H1299,H460,Huh-7and 2clinical samples were tested by using established method.Results An optimized sequencing system,consisted of P2-1[with 0% dimethyl sulphoxide（DMSO）,using slow-down PCR conditions]and P1-2（with 1% DMSO,using normal PCR conditions）,was successfully used in detection of human cell line H1299,H460,Huh-7and 2clinical samples,which could be applied for sequencing the CpG islands located at promoter region of TCRP1 gene.Conclusion A system for sequencing the CpG islands located at promoter region of TCRP1 gene was successfully established.