卵巢早衰患者卵泡刺激素受体基因突变的初步研究

对１４例受试者的卵泡刺激素（ＦＳＨ）受体基因进行了测试。１４例包括４例正常对照；１个有４个同胞姊妹的家庭，其中２人为卵巢早衰（ＰＯＦ）患者；其余５例为ＰＯＦ病人，１例为先天性卵巢不发育患者。采用限制片段长度多态性（ＲＦＬＰ），聚合酶链反应（ＰＣＲ），单链构象多态性（ＳＳＣＰ）和基因测序方法对受试者ＦＳＨ受体基因的第１～４、９和１０外显子作了检测。ＲＦＬＰ分析提示一些非特异性的变异可见于１例正常对照以及其他个别受试者；ＰＣＲ法未能检出异常的基因片段；ＳＳＣＰ筛选出第１０Ｄ外显子片段恒定呈现三种类型的ＤＮＡ移动带；对其中１０例的片段基因测序，发现４例纯合子，３例杂合子的基因片段在９１９位置上的腺嘌呤（Ａ）改变为鸟嘌呤（Ｇ），氨基酸由苏氨酸变成丙氨酸，这一改变在患者和对照都可见，但反复实验证明与ＤＮＡ碱基序列的立体构形有明确相关，而其他变异则未见此现象。提示ＦＳＨ受体基因的外显子的变异在正常人群中是存在的。ＰＯＦ很可能是一种潜在的多基因遗传疾病，而非一个特定的碱基变异所至。

A preliminary study about the mutations in the FSH receptor genes of females with premature ovarian failure and controls

Objective: To search for possible mutations or changes in FSH receptor gene structure unique to the premature ovarian failure (POF) patients. Methods: Forteen subjects,including 4 normally menstration controls,a familial with 4 sibling sisters, two of them developed POF, 5 POF patients and one female with 46, XX gonadal dysgenesis (GD). The restriction fragment length polymorphism(RFLP) analysis, polymerase chain reaction(PCR), single strand conformational polymorphism (SSCPs) and sequencing analysis were used to screen their FSH receptor gene on exon 1￣4,9 and 10. Results: RFLP did not reveal any insertions,delations or mutations relating to POF.However,another control subject displayed polymorphisms with Hinf I, Mbo I and Nco I analysis. No significant results were detectable by agarose gel electrophoretic analysis of PCR products.SSCPs analysis revealed a polymorphisms difference in exon 10 of the FSH receptor gene among both controls and POF subjects,with some subjects being homozygous and others being heterozygous. Sequencing revealed sporadic mutations in 1 RFLP positive control. Another polymorphism observed in 6 of 10 subjects including controls and POF,had A changed to G at position 919 which caused Thr 307 to be changed to Gly. This change caused a band shift in SSCPs. Conclusions: The point mutations in the FSH receptor gene occur commonly. The polymorphism G919 may be a key determinant of DNA conformation.The next work is to detect functional mutation among more patients and families.