牛卵巢黄体细胞增殖和孕酮分泌相关基因的筛选及其表达研究  

Screening and Expression Analysis of Genes Related to Proliferation and Progesterone Secretion of Luteal Cells in Bovine Ovary

作　　者：赵园园 孟金柱 伍春亚 潘春威 ZHAO Yuanyuan;MENG Jinzhu;WU Chunya;PAN Chunwei(Guizhou Provincial Key Laboratory for Biodiversity Conservation and Utilization in the Fanjing Mountain Region,Tongren University,Tongren 554300,China)

机构地区：[1]铜仁学院贵州省梵净山地区生物多样性保护与利用重点实验室,贵州铜仁554300

出　　处：《河南农业科学》2022年第10期125-133,共9页Journal of Henan Agricultural Sciences

基　　金：贵州省教育厅青年人才成长项目(黔教合KY字[2022]068);铜仁市科技计划项目(铜市科研[2020]80号);贵州省重点实验室项目(黔科合平台人才[2020]2003号);铜仁市科技局科技支撑计划项目(铜市科研[2020]128号)。

摘　　要：旨在筛选牛卵巢大黄体细胞(LLC,直径>25μm)和小黄体细胞(SLC,直径12~25μm)之间的差异表达基因,获得黄体细胞增殖和孕酮分泌的特征性基因。使用R软件limma包对GSE83524芯片数据进行分析,筛选出LLC和SLC中的差异表达基因;使用DAVID软件对差异基因进行GO功能注释及KEGG信号通路分析;利用在线软件String和Cytoscape构建蛋白质-蛋白质相互作用(PPI)网络并将其可视化;利用软件Cytoscape中的Cyto-Hubba插件筛选其中节点度最高的10个关键(Hub)基因;采用实时荧光定量PCR方法对获得的黄体细胞增殖和孕酮分泌的特征性基因在思南黄牛上进行验证。结果表明,在牛卵巢LLC和SLC中共筛选出226个差异表达基因;GO功能注释结果显示,226个差异表达基因注释到3个类别,共39组;KEGG分析结果表明,显著富集的通路有25条,其中PI3K-Akt信号通路与黄体细胞增殖和孕酮分泌密切相关;PPI网络分析获得10个节点度最高的Hub基因;实时荧光定量PCR检测结果表明,VWF、CYP19A1、CYP17A1、PRLR、BMPR2在LLC和SLC中的表达趋势与GEO芯片数据结果一致。The aim is to screen out the differentially expressed genes between large luteal cells(LLC,with the diameter>30μm)and small luteal cells(SLC,with the diameter of 12—25μm),and to obtain the characteristic genes related to proliferation and progesterone secretion of luteal cells.The GSE83524microarray data were analyzed to screen out the differentially expressed genes using the limma package of R software.GO functional annotation and KEGG signaling pathway analysis of differentially expressed genes were performed by DAVID.The online softwares String and Cytoscape were used to construct and visualize the protein-protein interaction(PPI)network.The Cyto-Hubba in Cytoscape was used to screen10 Hub genes with the highest node degree.The characteristic genes of luteal cell proliferation and progesterone secretion were verified by real-time PCR on Sinan yellow cattle.The results showed that a total of 226 differentially expressed genes were screened in LLC and SLC of bovine ovary.GO functional annotation results showed that 226 differentially expressed genes were annotated into 3 categories,a total of 39 groups.KEGG analysis showed that 25 pathways were significantly enriched,among which PI3K-Akt signaling pathway was closely related to luteal cell proliferation and progesterone secretion.PPI network interaction analysis obtained 10 Hub genes with the highest node degree.The results of real-time PCR showed that the expression trends of VWF,CYP19A1,CYP17A1,PRLR and BMPR2 in LLC and SLC were consistent with the results of GEO chip data.

关 键 词：思南黄牛 PI3K-AKT信号通路 小黄体细胞 大黄体细胞 孕酮 

分 类 号：S823[农业科学—畜牧学]