香猪卵巢circ_CSPP1的鉴定及功能预测分析

Identification and Function Prediction Analysis of circ_CSPP1 in Ovarian Tissues of Xiang Pigs

研究旨在揭示香猪卵巢组织中circ_CSPP1的潜在功能。利用CIRI-full拼接circ_CSPP1的序列,circBase数据库在线blat分析circ_CSPP1的保守性,采用RT-PCR和Sanger测序技术从香猪卵巢分离和鉴定circ_CSPP1,采用miRanda、RNAhybrid和PITA软件预测和分析circ_CSPP1结合的miRNA分子及miRNA的靶基因,对靶基因进行GO和KEGG富集分析。结果显示,分离到的circ_CSPP1是保守的,由CSPP1基因的外显子8~12组成,可作为ssc-miR-324、ssc-miR-10a-5p、ssc-miR-9847-3p、ssc-miR-365-5p、ssc-miR-92b-5p、ssc-miR-885-3p、ssc-miR-7139-3p、ssc-miR-9851-3p和ssc-miR-33b-3p等14个miRNAs的分子海绵,靶向755个蛋白编码基因。GO富集分析表明,circ_CSPP1的靶基因显著富集到153个GO条目,其中细胞成熟、细胞分裂位点、细胞迁移的负调控、Wnt信号通路、细胞周期蛋白结合、子宫发育、MAPK活性的激活、黏着斑和卵裂沟等条目与卵巢功能相关。KEGG富集分析表明,circ_CSPP1的靶基因富集到260个信号通路,其中显著富集的有29个信号通路,主要富集于癌症通路、细胞周期、雌激素信号通路、PI3K-AKT信号通路、卵巢类固醇生成、催产素信号通路和细胞凋亡等信号通路。综上,circ_CSPP1可作为许多miRNA的分子海绵调控香猪卵巢颗粒细胞的增殖与凋亡、卵泡成熟、排卵过程、卵巢类固醇生成和卵巢早衰等生物学过程。

The purpose of this study was to explore the potential functions of circ_CSPP1 in the ovarian tissues of Xiang pigs.In this study,the sequence of circ_CSPP1 was spliced by CIRI-full,the conservative analysis of circ_CSPP1 was calculated by online blat of circBase database,and the characteristics of circ_CSPP1 in the ovarian tissues of Xiang pigs were identified and analyzed by RT-PCR technology and Sanger sequencing technology.Moreover,miRanda,RNAhybrid and PITA softwares were used to predict and analyze the circ_CSPP1 bound miRNA molecular and miRNA target genes.Then GO and KEGG enrichment analyses were performed for the target genes.The results showed that the isolated circ_CSPP1 was conservative and originated from exons 8-12 of CSPP1 gene,which could be served as molecular sponges of 14 miRNAs,such as ssc-miR-324,ssc-miR-10a-5p,ssc-miR-9847-3p,ssc-miR-365-5p,ssc-miR-92b-5p,ssc-miR-885-3p,ssc-miR-7139-3p,ssc-miR-9851-3p,ssc-miR-33b-3p etc,and targeted 755 protein genes.GO enrichment analysis showed that the target genes of circ_CSPP1 were significantly enriched in 153 GO terms,in which cell maturation,cell division sites,negative regulation of cell migration,Wnt signaling pathway,cyclin binding,uterine development,activation of MAPK activity,focal adhesion and cleavage groove were related to ovarian function.KEGG enrichment analysis showed that the target genes of circ_CSPP1 were enriched into 260 signal pathways,of which 29 signal pathways were significantly enriched,and the target genes mainly concentrated in cancer pathway,cell cycle,estrogen signaling pathway,PI3K-AKT signaling pathway,ovarian steroidogenesis,oxytocin pathway and apoptosis etc.In conclusion,circ_CSPP1 might act as molecular sponges of many miRNAs to regulate the biological processes including proliferation and apoptosis of ovarian granulosa cells,follicle maturation,ovulation process,ovarian steroidogenesis and premature ovarian failure in the ovarian tissues of Xiang pigs.