Ghadami M, Salama S A, Khatoon N, Chilvers R, Nagamani M, Chedrese P J, Al-Hendy A
Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, TX 77555, USA.
Mol Hum Reprod. 2008 Jan;14(1):9-15. doi: 10.1093/molehr/gam077. Epub 2007 Dec 14.
Resistance ovarian syndrome is a heterogeneous disorder inherited as a Mendelian recessive trait and characterized by infertility, primary amenorrhea, normal karyotype and elevated serum FSH and LH levels. An inactivating mutation, C566T, in FSH receptor gene (FSHR) has been identified initially in Finland. We investigated if an adenovirus expressing a normal copy of human FSHR (Ad-hFSHR) has the ability to: (i) transfect granulosa cell lines, (ii) render the transfected cell lines responsive to FSH stimulation and (iii) transcomplement the malfunctioning form of human FSHR gene with C566T mutation. COS-7, JC-410, JC-410-P450-scc-luc and JC-410-StAR-luc cell lines were infected by Ad-hFSHR followed by treatment with FSH. Functional activity of the Ad-hFSHR was tested by measuring cyclic adenosine monophosphate (cAMP) or luciferase activity in response to FSH stimulation, and showed 2-4.6-fold increases in Ad-hFSHR transfected cells compared with untransfected or Ad-LacZ transfected cells, indicating that Ad-hFSHR is functionally active and expressing hFSHR. Generation of cAMP in cells expressing only mutated hFSHR-T566 showed minimal increase after FSH stimulation. Co-transfection of Ad-hFSHR in these cells carrying the malfunction form of human FSHR caused significant increases of 2.2-7.4-fold in FSH dependent cAMP generation (P = 0.0007). We concluded that adenovirus expressing a normal human FSHR can compensate the inactivating human FSHR-C566T mutation and restore FSH responsiveness.
抗性卵巢综合征是一种遗传性异质性疾病，以孟德尔隐性性状遗传，其特征为不孕、原发性闭经、正常核型以及血清促卵泡激素（FSH）和促黄体生成素（LH）水平升高。最初在芬兰发现了促卵泡激素受体基因（FSHR）中的一种失活突变C566T。我们研究了表达人FSHR正常拷贝的腺病毒（Ad-hFSHR）是否有能力：（i）转染颗粒细胞系；（ii）使转染后的细胞系对FSH刺激产生反应；（iii）对具有C566T突变的人FSHR基因的功能异常形式进行反式互补。用Ad-hFSHR感染COS-7、JC-410、JC-410-P450-scc-luc和JC-410-StAR-luc细胞系，随后用FSH处理。通过测量对FSH刺激的反应中环磷酸腺苷（cAMP）或荧光素酶活性来测试Ad-hFSHR的功能活性，结果显示，与未转染或Ad-LacZ转染的细胞相比，Ad-hFSHR转染细胞中的活性增加了2至4.6倍，这表明Ad-hFSHR具有功能活性且表达hFSHR。仅表达突变型hFSHR-T566的细胞在FSH刺激后cAMP的生成仅有极小的增加。在携带人FSHR功能异常形式的这些细胞中共转染Ad-hFSHR，导致FSH依赖性cAMP生成显著增加2.2至7.4倍（P = 0.0007）。我们得出结论，表达正常人FSHR的腺病毒可以补偿失活的人FSHR-C566T突变并恢复FSH反应性。
