用PCR-SSCP分析检测睾丸女性化综合征患者雄激素受体基因突变

PCR SSCP ANALYSIS OF ANDROGEN RECEPTOR GENE MUTATIONS IN PATIENTS WITH TESTICULAR FEMINIZATION SYNDROMES

为进一步探讨睾丸女性化（Ｔｆｍ）综合征患者发病的分子机理，并为研究雄激素受体（ＡＲ）结构与功能关系提供资料，应用银染聚合酶链式反应－单链构象多态性（ＰＣＲ－ＳＳＣＰ）分析方法对４例Ｔｆｍ患者的ＡＲ基因８个外显子中的７个（Ｂ～Ｈ）分别进行了研究。结果表明，所有患者ＡＲ基因的上述外显子经ＰＣＲ扩增后均出现与外显子相应长度一致的条带，表明没有明显的缺失或插入突变。２例患者外显子Ｇ片段在行ＳＳＣＰ分析时分别有泳动变位，经ＤＮＡ双链循环测序证实两外显子各有一点突变（Ｖａｌ８６６Ｍｅｔ，Ｔ２９１９→Δ），这些突变均位于ＡＲ的雄激素结合区内。其中的缺失突变为国际上尚未报道过的新突变。研究表明，ＰＣＲ－ＳＳＣＰ分析是检测ＡＲ基因突变的快速、简便、可靠的方法。

In this study, single strand conformation polymorphism ( PCR SSCP) analysis was established to screen for androgen receptor (AR) gene mutation. The exons B￣H of AR gene of 4 patients with complete testicular feminization(Tfm) syndromes were analyzed using a combination of PCR SSCP and direct DNA sequencing. Two mutations in the AR gene which caused amino acid substitutions of AR (Va1866Met, T1919→△) respectively were identified in two patients. No mutations in exons B￣H have been found in the other two patients. This analysis has provided valuable information for studying the molecular mechanism of Tfm.