多囊卵巢综合征患者与正常排卵妇女GV期卵母细胞转录组的比较

doi:10.13481/j.1671-587x.20180321

吉林大学学报(医学版) ›› 2018, Vol. 44 ›› Issue (03): 568-573.doi: 10.13481/j.1671-587x.20180321

多囊卵巢综合征患者与正常排卵妇女GV期卵母细胞转录组的比较

杜琛, 陈秀娟, 侯石磊, 赵杰

内蒙古医科大学附属医院生殖医学中心, 内蒙古呼和浩特 010050

收稿日期:2017-08-08 出版日期:2018-05-28 发布日期:2018-05-31
通讯作者: 陈秀娟,教授,硕士研究生导师(Tel:0471-3451651,E-mail:90098687@sina.com) E-mail:90098687@sina.com
作者简介:杜琛(1986-),女,内蒙古自治区呼和浩特市人,助理研究员,遗传学博士,主要从事胚胎发育方面的研究。
基金资助:
内蒙古自治区科技厅自然科学基金资助课题（2015BS0802）

Comparsion of GV oocytes transcriptome between PCOS patients and normal ovulatory women

DU Chen, CHEN Xiujuan, HOU Shilei, ZHAO Jie

Reproductive Medicine Center, Affiliated Hospital, Inner Mongolia Medical University, Hohhot 010050, China

Received:2017-08-08 Online:2018-05-28 Published:2018-05-31

摘要/Abstract

摘要： 目的：探讨控制性卵巢刺激（COS）对多囊卵巢综合征（PCOS）患者和正常排卵妇女GV期卵母细胞中差异表达基因和主要信号转导通路的影响，从而筛选出影响PCOS患者卵母细胞发育的关键基因。方法：选择接受GnRH-a长方案将调的控制性超排卵的PCOS患者3例（PCOS组）和同期因男性不孕因素接受长方案将调的正常排卵妇女3人（对照组），通过酶消化法分离颗粒细胞，收集经过卵胞浆内单精子显微注射（ICSI）之后废弃的未成熟卵母细胞。构建cDNA文库，在Illumina MiSeq测序平台进行测序，并通过RT-PCR技术对测序数据进行体外验证（每组3个重复）。结果：获得了510 024 82个序列读取片段（reads），包含8 G碱基序列信息。通过生物信息学软件共找到63个差异表达基因，极显著上调的基因19个，极显著下调基因有44个（Fold Change>4，FDR<0.01）。PCOS组患者血管内皮生长因子（VEGF）和脂肪酸脱氢酶1（FADS1） mRNA表达水平明显高于对照组（P<0.05），Runt相关转录因子2（RUNX2）、趋化因子1（CXCL1）和热体克蛋白27（Hsp27） mRNA表达水平明显低于对照组（P<0.05），与功能验证结果一致。差异基因基于GO功能注释可分为磷酰化、细胞自噬和转录调控等多个分支，利用KEGG数据库作为参考，这些基因主要富集于磷脂酰肌醇-3-激酶/蛋白激酶B（PI3K-Akt）、转化生长因子β（TGF-β）、Hippo、p53和过氧化物酶体增殖剂激活受体（PPAR）信号通路中。结论：PCOS患者GV期卵母细胞中差异表达的基因可能影响卵泡发育和卵母细胞成熟，从而导致胚胎质量低下。

关键词: 多囊卵巢综合征, 高通量测序, 卵母细胞, 控制性卵巢刺激

Abstract: Objective: To investigate the effects of controlled ovarian stimulation(COS) on the differentially expressed genes in GV oocytes and main signal transduction pathways in the polycystic ovary syndrome (PCOS) patients and the normal ovulatory women,and to screen the key genes impacting the development of oocytes of the PCOS patients. Methods: During controll ovarian hyperstimulation with GnRH-a long protocol, 3 patients with PCOS(PCOS group) and 3 normal ovulatory women due to male infertility factor (control group)were selected.Enzyme digestion was used to islolate the granule cells.The dumped immature oocytes were collected after intracytoplasmic sperm injection(ICSI).The cDNA library was constructed and sequencing was performed in Illumina MiSeq sequencing platform and RT-PCR was used to confirm the data obtained in vivo. Results: A total of 510 024 82 sequence reads were obtained,and 8 G base sequence information were contained.A total of 63 differentially expressed genes were found by bio-informatics software,including 19 significant up-regulation genes and 44 significant down-regulation genes (Fold Change>4,FDR<0.01).The expression levels of vascular endothelial growth(VEGF) and fatly acid dehydrogenase 1(FADS1) mRNA in the patients in PCOS group were significantly higher than those in control groups(P<0.05),and the expression levels of Runt-related transcription factor 2(RUNX2),chemokine factor 1 (CXCL1) and heat shock protein27(Hsp27) mRNA were significantly lower than those in control group (P<0.05).These above results were consistent with the functional verification results.According to the GO function annotation,these genes were divided into several branches,including phosphorylation,autophagy,and transcriptional regulation,etc.Referring to the KEGG database,the differential genes were mainly associated with phosphatidylinositol -3-kinase (PI3K-Akt),transforming growth factor β(TGF-β) Hippo,p53 and peroxisome proliferators-activated receptor(PPAR) signal pathways. Conclusion: The differentially expressed genes in GV oocytes of the PCOS patients may affect the development of follicle and maturation of oocytes and lead to poor quality of embryo.

Key words: polycystic ovary syndrome, controlled ovarian stimulation, oocytes, high-throughput sequencing

中图分类号:

Q344.13

杜琛, 陈秀娟, 侯石磊, 赵杰. 多囊卵巢综合征患者与正常排卵妇女GV期卵母细胞转录组的比较[J]. 吉林大学学报(医学版), 2018, 44(03): 568-573.

DU Chen, CHEN Xiujuan, HOU Shilei, ZHAO Jie. Comparsion of GV oocytes transcriptome between PCOS patients and normal ovulatory women[J]. Journal of Jilin University Medicine Edition, 2018, 44(03): 568-573.

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多囊卵巢综合征患者与正常排卵妇女GV期卵母细胞转录组的比较

Comparsion of GV oocytes transcriptome between PCOS patients and normal ovulatory women

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