摘要
生长停滞特异性蛋白6(growth arrest specific protein 6,GAS6)在肿瘤发生发展中发挥重要作用,其信号转导参与细胞增殖、黏附与迁移,但它在子宫内膜异位症(endometriosis, EMs)的相关功能及分子机制尚不明确。本研究从GEO数据库检索并下载子宫内膜异位症相关转录物组数据集,并对其进行GEO在线分析,筛选差异表达基因并进行GO聚类和KEGG通路富集分析。利用10例无内异症且无明确疾病妇女的在位子宫内膜,以及11例卵巢巧克力囊肿病人异位子宫内膜,对3个以上数据集共有的差异基因的mRNA水平进行实时荧光定量PCR验证。在子宫内膜异位症临床样本中,采用免疫组化、实时荧光定量PCR验证关键调控因子GAS6及上皮间充质转化(epithelial mesenchymal transition, EMT)标记基因的表达水平,并利用免疫荧光对GAS6和E-钙黏着蛋白(E-cadherin)进行共标。研究发现:从4个转录物组数据集中共筛选出47个差异表达基因,其主要富集于细胞迁移等过程以及MAPK、PI3K-AKT、紧密连接等相关信号通路。3个以上数据集所共有的9个差异基因在子宫内膜异位症病人中的mRNA水平均符合生物信息学分析的结果。GAS6在子宫内膜异位症病人异位内膜中的表达水平高于对照组(P<0.05),并且子宫内膜异位症病人的内膜组织中存在EMT现象,EMT的标志物E-钙黏着蛋白表达水平下调(P<0.05)、波形蛋白(vimentin)表达水平上调(P<0.01)。在GAS6高表达的子宫内膜异位症患者异位子宫内膜腺上皮细胞中,E-cadherin显示低表达,提示GAS6可能在子宫内膜异位症中介导EMT过程。综上所述,本研究初步揭示GAS6在子宫内膜异位症病人中高表达,及其可能介导EMT过程参与子宫内膜异位症的发生与发展,为子宫内膜异位症的临床治疗提供潜在靶标。
Growth arrest specific protein 6(GAS6) plays an important role in the occurrence and development of tumors, and its signal transduction is involved in cell proliferation, adhesion and migration, but its related functions and molecular mechanisms in endometriosis(EMs) are still unclear. In this study, we searched and downloaded the transcriptome datasets of EMs from GEO database and performed GEO online analysis, and then screened out the differentially expressed genes and performed cluster analysis based on GO and KEGG pathway. The mRNA levels of the differentially expressed genes shared by more than three datasets were verified by qRT-PCR in the endometrium of ten women with no endometriosis and no clear disease and the ectopic endometrium of 11 patients with ovarian chocolate cysts. Immunohistochemistry and qRT-PCR were used to verify the expression of GAS6 and epithelial mesenchymal transition(EMT) marker genes, and immunofluorescence was used to co-label GAS6 and E-cadherin in endometriosis clinical samples. In this study, a total of 47 differentially expressed genes were screened out of the four transcriptome datasets, which were mainly enriched in processes such as cell migration and related signal pathways such as MAPK, PI3 K-AKT, and tight junction. The mRNA levels of the nine differentially expressed genes shared by more than three datasets in endometriosis patients were consistent with the results of bioinformatics analysis. GAS6 expression levels in ectopic endometrium of EMs patients are higher than the control group(P < 0.05), and EMs patients have the characteristics of EMT in the ectopic endometrial tissue, that is, the expression of E-cadherin is down-regulated(P < 0.05) and the expression of vimentin is up-regulated(P < 0.01). The expression of E-cadherin in the ectopic endometrial glandular epithelial cells of EMs patients is low while the expression of GAS6 is up-regulated, suggesting that GAS6 may mediate the EMT process in endometriosis. In conclusion, this study reveals that GAS6 is highly e
作者
刘畅
刘太行
何帆
池余刚
杨博萍
肖仕全
丁裕斌
李方方
LIU Chang;LIU Tai-Hang;HE Fan;CHI Yu-Gang;YANG Bo-Ping;XIAO Shi-Quan;DING Yu-Bin;LI Fang-Fang(Joint International Research Laboratory of Reproduction&Development,School of Public Health and Management,Chongqing Medical University,Chongqing 400016,China;Reproductive Center of the Second Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China;Department of Gynecology,Chongqing Maternal and Child Health Hospital,Chongqing 400016,China;Reproductive Medicine Center of Third Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China)
出处
《中国生物化学与分子生物学报》
CAS
CSCD
北大核心
2021年第7期917-926,共10页
Chinese Journal of Biochemistry and Molecular Biology
基金
重庆市渝中区基础研究与前沿探索项目(No.20190134)资助。
关键词
子宫内膜异位症
差异基因
生长停滞特异性蛋白6
上皮间充质转化
endometriosis(EMs)
differential genes
growth arrest specific protein 6(GAS6)
epithelial mesenchymal transition(EMT)
