Research Progress on Mutations and Single Nucleotide Polymorphisms of Follicle Stimulating Hormone Receptor Gene

卵泡刺激素受体(FSHR)是由FSHR基因编码的G蛋白耦联受体蛋白,由胞外区、跨膜区及胞内区3部分构成.胞外区与FSH特异性结合组成FSH/FSHR系统,在人类生殖过程中发挥着重要作用.FSHR基因上突变基因分为活性突变和失活突变2种,失活突变可能导致原发或继发性闭经、高促性腺激素性功能障碍、卵巢早衰及生精功能障碍等生殖疾病,活性突变主要与卵巢过度刺激综合征(OHSS)关系密切.FSHR基因的点突变出现概率非常小,大部分仅有1次报道,而大多数FSHR基因突变为单核苷酸多态性.卵巢和睾丸的正常发育及发挥功能均依赖于完整的FSHR介导,FSHR突变对两性生殖表型的影响存在着差异.     

FSH受体基因多态性在中国IVF人群中分布及与卵巢反应性关系的研究

目的:探讨IVF-ET/ICSI患者中FSH受体(FSHR)基因多态性对控制促排卵中卵巢反应性及妊娠结局的影响。方法:分析行IVF-ET/ICSI的223例患者资料,其中长方案174例,短方案37例,微刺激方案12例。通过PCR扩增、纯化、基因测序分别测得223例患者FSHR体基因在外显子10上第680位及307位的基因型,根据不同基因型分为Asn680/Asn680、Ser680/Ser680、Asn680/Ser680;Thr307/Thr307、Ala307/Ala307、Ala307/Thr307各3组。比较223例患者不孕因素在各组的分布;比较174例长方案促排卵患者不同FSHR基因型亚组的卵巢反应及妊娠结局。结果:223例患者中,FSHR基因680位点上Asn/Asn组占52.0%,Ser/Ser组占12.1%,Asn/Ser组占35.9%,307位点上Ala/Ala组占12.1%,Thr/Thr组占48.4%,Ala/Thr组占39.5%。174例长方案患者中Ser680/Ser680组及Ala307/Ala307亚组的基础FSH值显著高于其它2个基因型组(P<0.05);FSH受体基因680、307位点多态性的改变在OHSS发生上无预测作用(P>0.05),但OHSS发生组中SS680的分布频率(4.6%)明显低于非OHSS组(15.1%)。FSHR基因680、307位点多态性的改变与临床妊娠率无相关性。结论:FSHR基因680、307位多态性的分布存在差异,FSHR基因多态性Ser680和AA307可能与卵巢反应性有关,FSHR基因680、307位多态性与临床妊娠率无关。     

卵泡刺激素受体与卵巢癌

卵巢癌是女性生殖系统最常见恶性肿瘤之一,以高复发率、高死亡率为特点.卵巢癌病因至今尚不明.大量研究显示,卵泡刺激素(FSH)与卵巢癌的发生密切相关.FSH对卵巢的作用主要是通过与卵泡刺激素受体(FSH Receptor,FSHR)特异性结合而介导.近年研究发现,在FSHR基因第10外显子及启动子序列的单核苷酸多态性,以及外显子不同剪接机制所致的异构体,可能对FSHR表达及其与FSH的结合产生影响.而FSHR缺失或过度表达都可能影响正常的信号转导,从而促进肿瘤发生.     

卵泡刺激素受体基因与多囊卵巢综合征关系的初步研究

目的:探讨卵泡刺激素受体基因第7、10(A)外显子基因突变与多囊卵巢综合征(PCOS)发病的关系。 方法:提取56例PCOS患者及38例对照者的外周血基因组DNA,采用PCR及RFLP(限制性片段长度多态性)方法研究FSHR第7、10(A)外显子基因多态性。 结果:PCOS组FSHR第7外显子49/49型96.43%,86/49型3.57%,第10(A)外显子128/128型98.21%,128/216型1.79%,与对照组相比较,其基因型分布差异无显著性(P>0.05)。PCOS组第7外显子49型等位基因频率为98.21%,86型等位基因频率为1.79%,第10(A)外显子128型等位基因频率为99.11%,216型等位基因频率为0.89%。结论:中国人群存在以上两位点突变,但突变率在PCOS组及对照组不存在差异。PCOS组LH /FSH比值、高雄激素血症与等位基因86型、216型存在一定的相关趋势。     

卵巢颗粒细胞中卵泡刺激素的受体后信号传导通路

卵泡刺激索(FSH)是下丘脑-垂体-性腺轴中的关键激素,通过与颗粒细胞的FSH受体(FSHR)结合,发挥卵泡募集、促卵泡生长和卵母细胞成熟的作用,并与黄体生成素(LH)一起,介导排卵过程,促进和维持正常的性腺发育和生殖功能.研究颗粒细胞中FSH与FSHR结合后的信号传导通路(即受体后信号传导通路)机制,对了解颗粒细胞在卵泡发育过程中的作用以及对治疗不孕、不育药物的研制具有重要意义.本文将对颗粒细胞中FSH的受体后信号传导及功能调节的分子学机制的研究进展做一综述.     

卵泡刺激素受体与卵巢癌

卵巢癌是女性生殖系统最常见恶性肿瘤之一,以高复发率、高死亡率为特点。卵巢癌病因至今尚不明。大量研究显示,卵泡刺激素（FSH）与卵巢癌的发生密切相关。FSH对卵巢的作用主要是通过与卵泡刺激素受体（FSH Receptor,FSHR）特异性结合而介导。近年研究发现,在FSHR基因第10外显子及启动子序列的单核苷酸多态性,以及外显子不同剪接机制所致的异构体,可能对FSHR表达及其与FSH的结合产生影响。 而FSHR缺失或过度表达都可能影响正常的信号转导,从而促进肿瘤发生。     

长三角地区重度少弱精子症与卵泡刺激素受体基因单核苷酸多态性相关性研究

目的:研究卵泡刺激素受体(FSHR)基因Thr307Ala(rs6165)和Asn680Ser(rs6166)单核苷酸多态性(SNP)基因型分布情况及其与中国长三角地区重度少弱精子症的关联性。方法:外周血提纯DNA,PCR扩增后直接测序分析200名已育男性(已生育组)和150名重度少弱精子症不育男性(不育组)FSHR基因Thr307Ala和Asn680Ser位点的SNP,并用χ2检验进行相关性分析。结果:FSHR基因Thr307Ala和Asn680Ser多态性位点的基因型分布在已生育组和不育组间无统计学差异(P>0.05)。Thr-Asn/Ala-Asn和Thr-Ser/Thr-Ser双倍型在已育男性与重度少弱精子症患者间分布差异有统计学意义(P<0.05),组间FSHR的单倍型Thr-Asn和Ala-Asn之间亦有统计学差异(P<0.05)。结论:FSHR基因Thr307Ala和Asn680Ser的2个多态性位点特定的单倍型和双倍型与男性不育有一定的相关性。     

Notch信号通路在小鼠卵巢皮层组织的表达变化

目的:探索Notch信号通路在调控卵巢生殖干细胞增殖和衰老中的作用。方法:分别提取3日龄、2月龄、12月龄及20月龄小鼠卵巢总RNA,q PCR法和免疫组织化学法分别检测Notch通路信号分子受体Notch1及靶基因Hes1、Hes5,生殖细胞标志基因Mvh、胚胎干细胞标志物Oct4的m RNA及蛋白的表达情况。双免疫荧光检测卵巢皮质层中Mvh和Notch1的共表达情况。结果:Notch1、Hes1、Hes5、Mvh及Oct4这5个基因的m RNA表达水平随着小鼠生殖年龄增长呈现由高到低的一致性动态变化;Notch1、Jagged1(Notch1配体)及Hes1的蛋白表达水平随着鼠龄增长逐渐减弱;双免疫荧光结果显示Mvh和Notch1共表达趋势随着鼠龄增长而减弱。结论:Notch信号通路活性在小鼠卵巢组织及卵巢上皮组织中的表达随着鼠龄的增长而逐渐减弱,与对应的生殖干细胞标志基因的表达水平呈现一致性,Notch信号通路参与卵巢生殖干细胞增殖的调控。     

超排卵周期卵巢的反应性与卵泡颗粒细胞中卵泡刺激素受体表达的关系

目的探讨超排卵周期卵巢的反应性与颗粒细胞中卵泡刺激素受体(FSHR)表达的关系。方法采用蛋白印迹法,测定因输卵管性不孕或者男性因素不孕的60例不孕症患者的卵泡颗粒细胞FSHR蛋白的水平;采用电化学发光法测定血清雌二醇峰值。根据超排卵周期中发育卵泡数,将60例患者分为卵巢低反应组(20例)、卵巢中反应组(20例)及卵巢高反应组(20例),比较各组FSHR蛋白表达水平,并分析FSHR表达水平与卵泡数、雌二醇峰值的相关性。结果(1)卵巢低反应组、卵巢中反应组及卵巢高反应组的FSHR蛋白表达分别为0·19±0·07、0·34±0·16及0·45±0·18,3组比较,差异均有统计学意义(P<0·001);卵泡数分别为(2·5±0·5)、(7·9±1·9)及(21·6±3·8)个,3组比较,差异均有统计学意义(P<0·01);雌二醇峰值分别为3441、7864及22486pmol/L,3组比较,差异有统计学意义(P<0·01)。(2)3组患者超排卵周期颗粒细胞FSHR表达水平与卵泡数及雌二醇峰值均呈显著正相关关系(rs=0·52及0·71,P<0·01)。结论超排卵周期中,卵巢的反应性与颗粒细胞FSHR蛋白表达有关,FSHR表达水平的高低反映不同的卵巢反应类型。     

不同卵巢储备患者促性腺激素受体表达及影响因素分析

目的探讨不同卵巢储备女性颗粒细胞促卵泡激素受体(FSHR)、促黄体生成素受体(LHR)mRNA相对表达水平与获卵情况的关系,以及与其他促排卵治疗过程中相关指标的影响。方法收集850例患者卵泡液及颗粒细胞。纳入患者分为卵巢功能正常组(n=419)、预期低反应组(n=255)、多囊卵巢综合征(PCOS)组(n=176)。采用反转录-实时定量PCR(qRT-PCR)法测定FSHR、LHR、抗苗勒管激素(AMH)及AMHⅡ型受体(AMHRⅡ) mRNA的表达,分析以上指标在不同卵巢储备患者中表达差异及影响因素。结果 (1) 3组患者在年龄、体质量指数(BMI)、窦卵泡数(AFC)、基础激素水平、用药情况、FSHR mRNA及获卵数等方面差异均有统计学意义(P0.05)。(2) PCOS组促排卵后卵巢高反应患者FSHR mRNA表达量低于正常反应患者(P0.05)。(3)对卵巢功能正常组患者FSHR mRNA主要与AMHmRNA(r=0.404,P0.001)、LHR mRNA(r=0.388,P0.001)呈正相关,预期低反应组患者FSHR mRNA主要与LHR(r=0.415,P0.001)呈正相关,PCOS患者FSHR mRNA主要与AMHRⅡmRNA(r=0.311,P0.001)呈正相关。结论不同卵巢储备患者促性腺激素(Gn)受体表达量存在差异,且Gn受体与AMHmRNA表达具有显著相关性,可能与卵巢储备异常发病机制相关,临床Gn用药可参考Gn受体表达情况进行选择。     

Mutations in the follicle-stimulating hormone-beta (FSH beta) and FSH receptor genes in mice and humans

Follicle-stimulating hormone (FSH), a dimeric glycoprotein synthesized in the anterior pituitary gland, is important for the production of sex steroids and gametes. FSH-beta (FSH beta) and FSH receptor (FSHR) knockout mice display impaired ovarian follicular development and infertility in females and small testes, oligospermia, and fertility in males. Humans with FSH beta gene mutations tend to have a more severe phenotype than those with FSHR gene mutations, although infertility and varying degrees of impaired sex steroid production occur in both types of mutations. Data from human and mouse mutations in the FSH beta and FSHR genes suggest that FSH is necessary for normal pubertal development and fertility in males and females.     

Effect of follicle-stimulating hormone receptor Asn680Ser polymorphism on the outcomes of controlled ovarian hyperstimulation: an updated meta-analysis of 16 cohort studies

PurposeThe purpose of this study is to evaluate the influence of follicle-stimulating hormone receptor (FSHR) Asn680Ser polymorphism on the ovarian response to exogenous follicle-stimulating hormone (FSH) and clinical outcomes in women undergoing controlled ovarian hyperstimulation (COH).MethodsA database search was conducted to identify the eligible studies that investigated the effect of FSHR Asn680Ser polymorphism on ovarian response and clinical outcomes. A pooled analysis was performed with the odds ratio (OR) or weighted mean difference (WMD) and their respective 95 % confidence interval (CI) by the STATA software with random effects model.ResultsSixteen cohort studies comprising a total of 4287 subjects were included. The number of retrieved oocytes was significantly fewer in subjects with the SS genotype at position 680, compared to subjects with the NN or NS genotype (WMD = −1.36, 95 % CI = −1.85 to −0.87). Lack of association was detected between the genotypes (SS genotype vs. NN or NS genotype) and clinical outcomes such as exogenous FSH dose (WMD = 98.96 IU, 95 % CI = −22.33 to 220.24), poor response (OR = 1.08, 95 % CI = 0.71–1.64), ovarian hyperstimulation syndrome (OHSS) (OR = 1.58, 95 % CI = 0.41–6.07), and clinical pregnancy rate (OR = 1.10, 95 % CI = 0.86–1.40). However, poor ovarian response and number of retrieved oocytes were significantly influenced by the Asn680Ser polymorphism in the Asian subjects. In addition, no publication bias was detected.ConclusionFSHR Asn680Ser polymorphism might be a significant biomarker for predicting the number of retrieved oocytes and poor response, especially in Asian subjects. Other outcomes such as exogenous FSH dose, OHSS, and pregnancy rate were not influenced by FSHR Asn680Ser polymorphism.     

FSHR基因单核苷酸多态性与围绝经期妇女卵巢功能的相关性研究

目的探讨围绝经期妇女的FSHR基因单核苷酸多态性（SNPs）与卵巢功能衰退的相关性。方法根据血清FSH水平,分为卵巢功能正常组（正常组）84例、卵巢储备功能下降组（下降组）77例和卵巢功能衰竭组（衰竭组）34例。应用SNaPshot技术,进行FSHR基因Ser680Asn多态性检测。结果 FSHR基因Ser680Asn位点基因型频率分布三组之间差异有统计学意义（χ2=8.5648,P=0.0138）。两两分析,正常组和下降组之间差异有统计学意义（χ2=9.0386,P=0.0109）,正常组和衰竭组之间差异有统计学意义（χ2=8.3186,P=0.0156）。等位基因频率的分布在三组之间差异有统计学意义（χ2=7.6645,P=0.0056）。两两分析,正常组和衰竭组之间差异有统计学意义（χ2=8.3449,P=0.039）,下降组和衰竭组之间差异有统计学意义（χ2=3.8502,P=0.0497）。且卵巢功能随基因型的变化（Asn/Asn,Asn/Ser和Ser/Ser）呈现降低趋势（χ2=8.5648,P=0.0138）。结论 FSHR基因Ser680Asn单核苷酸多态性对围绝经期妇女卵巢功能有影响,与卵巢功能的下降和衰竭相关。     

Ovarian hyper-stimulation syndrome after spontaneous conception

Ovarian hyperstimulation syndrome (OHSS) is rather frequent (1–5%) in women submitted to superovulation with gonadotropins for in vitro fertilisation (IVF), whereas it is very rare in case of spontaneous ovulation. Spontaneous OHSS (sOHSS) was previously described to be associated to hydatiform mole, multiple conception, hypothyroidism in pregnancy. It may also depend on activating mutations of the FSH receptor (FSHR) gene that cause ovarian hyper-responsiveness to circulating FSH or even cross-responsiveness of FSHR to hormones having a structure similar to FSH, such as hCG or TSH. We report, herein, a case of sOHSS in a woman who conceived spontaneously. We checked the presence of all possible factors that could explain the onset of the syndrome, and we evidenced hypothyroidism and abnormally elevated hCG levels in the second trimester of pregnancy. The thorough molecular biology study of FSHR gene did not detect exonic mutations, but revealed the presence of intronic mutations whose role in the onset of sOHSS is still uncertain.     

Role of nerve growth factor and FSH receptor in epithelial ovarian cancer

The neurotrophins (NT) including nerve growth factor (NGF) are a family of related growth factors that are of major importance in the regulation of neuronal survival and differentiation. In the ovary, they can help in follicular maturation and ovulation by inducing the FSH receptor (FSHR). Current literature shows that perimenopausal ovarian surface epithelium (OSE) can also express FSHR. By G protein link, this FSHR is capable of precipitating neoplasia of OSE, which is the commonest in the ovary. NT are implicated as the cause of this aberrant expression of FSHR in OSE. By central action NT can lower serum FSH, as is found in ovarian cancer. Thus, NGF deregulates expression of FSHR in OSE and secretion of FSH from the pituitary. This phenomenon may hold the key to the hitherto unexplained carcinogenic process of sporadic epithelial ovarian cancer.     

Follicle-stimulating hormone polypeptide modified nanoparticle drug delivery system in the treatment of lymphatic metastasis during ovarian carcinoma therapy

Objective

Traditional chemotherapy drugs have an obvious drawback of nonspecific biodistribution in treating ovarian cancer. Follicle-stimulating hormone receptor (FSHR), a G-protein coupled receptor which is mainly expressed in reproductive system, is an important drug target in developing novel therapeutics.

Methods

Using a polypeptide of follicle-stimulating hormone (named as FSHP), a conjugated nanoparticle, FSHP-NP was developed to target FSHR in lymphatic metastasis of ovarian cancer. FSHP-NP was tested for recognition specificity and uptake efficiency on FSHR-expressing cells. A paclitaxel (PTX)-loaded FSHP-NP (FSHP-NP-PTX) was further developed and its anti-tumor effect was determined in vivo and in vitro.

Results

Taking NuTu-19 cells as an example, FSHP-NP-PTX displayed significantly stronger anti-cell proliferative and anti-tumor effects in a dose- and time-dependent manner when compared with free PTX or naked PTX-loaded nanoparticles (NP-PTX) in vitro. In vivo examinations showed that the size and weight of the lymph nodes were reduced in the FSHP-NP-PTX group.

Conclusion

FSHR as a novel therapeutic target in ovarian cancer and delivery of PTX via conjugated nanoparticle (FSHP-NP) might represent a new therapeutic approach in ovarian cancer.     

Inferring biallelism of two FSH receptor mutations associated with spontaneous ovarian hyperstimulation syndrome by evaluating FSH,LH and HCG cross-activity

Research question

What is the cumulative effect of two follicle-stimulating hormone receptor (FSHR) mutations in spontaneous ovarian hyperstimulation syndrome (sOHSS) pathogenesis? Are these mutations in the mono- or biallelic state?