喉鳞癌PTEN基因第5、8外显子的测序分析

Mutational Analysis of Exon 5,8 of the PTEN Gene in Laryngeal Squmaous Cell Carcinoma

目的探讨 PTEN 基因与喉鳞状细胞癌的关系。方法应用聚合酶链反应-单链构像多态性分析(polymerase chain reaction-single-strand conformation polymorphism,PCR-SSCP)和 TA 克隆 PCR 产物 DNA 测序技术,检测喉鳞状细胞癌新鲜或冰冻组织中抑癌基因 PTEN 第5、8外显子的突变情况。结果 60例喉鳞状细胞癌组织中使用 SSCP 检测发现7例显示电泳迁移率的改变。对这7例阳性样本的 PCR 产物进行 TA 克隆后测序,2例第5外显子第474位碱基发生 A→T的颠换。2例第5外显子第397、407位碱基发生 G→A的转换,导致错义突变;1例第5外显子第405位碱基发生 A→T的颠换。1例第8外显子第830位碱基发生 C→T的转换,导致错义突变;1例第8外显子第831位碱基发生 A→C颠换。结论 PTEN 基因的外显子5突变可能与喉癌的发生有一定的关系,其机制有待深入研究。

Purpose To detect mutations of exon 5 and 8 of the PTEN gene in laryngeal squmaous cell carcinoma (LSCC) and analyze the relationship between the mutation and ISCC.Methods Fresh tumor samples from 60 LSCC patients were examined using polymerase chain reaction-single-strand conformation polymorphism (PCR-SSCP) and TA cloning DNA sequencing method.Results An extra band in SSCP was shown in 7 of 60 samples.Exon 5 and 8 of PTEN were detected by TA cloning DNA sequencing method in the 7 cases.2 cases of exon 5,transversion GTF→GTA,and 2 cases of exon 5, transition GTA→ATA and TGT→TAT may result in missence mutation.1 case of exon 5,transversion ATA→ATT.and 1 case of exon 8,transition ACA→ATA may also result in missence mutation.1 case of exon 8 was transversion ACA→ACC. Conclusion The mutation of exon 5 of the PTEN gene may be related to the occurrence of LSCC.