香猪睾丸miR-449b和miR-222分子的发育性变化

Expressions of miR-449b and miR-222 in the testis of Xiang pigs at developmental stages

为了阐明miR-449b和miR-222对贵州从江香猪睾丸发育的调节作用。运用qPCR方法检测香猪睾丸组织中miR-449b和miR-222表达量的发育性变化,通过miRSystem数据库、Kobas数据库对预测的靶基因进行信号转导通路富集分析。经克隆测序,得到香猪miR-222序列,与猪的相似性为100%,而且物种之间miR-449b和miR-222两个分子的保守性都很高。香猪睾丸中miR-449b于1~4月龄期间的表达量逐渐上升,4月龄时表达量达到峰值,之后下降;miR-222的表达量呈现相反的变化趋势,于1~4月龄期间表达量逐渐下降,1月龄时最高,4月龄时最低,之后的表达量有所回升;已知物种的miR-449b和miR-222序列高度保守;经预测miR-449b靶基因有420个,富集于卵母细胞减数分裂、Notch信号通路、Ras信号通路等100个途径(P<0.05),miR-222的靶基因有429个,富集于细胞周期、Fox O信号通路、TGF-β信号通路等69个通路(P<0.05);miR-449b和miR-222的靶基因共享的通路为Wnt信号通路、MAPK信号通路、雌激素信号通路等42个(P<0.05)。

The present paper is aimed to explore the effects of miR-449 b and miR-222 on the testis development of Xiang pig. The expression patterns of miR-449 b and miR-222 molecules were determined using real-time quantity PCR method. Target genes of miR-449 b and miR-222 were deduced by miR System and Kobas databases,which were further applied to predict for the signal transduction pathway enrichment analysis. The sequence of pig miR-222 was first determined with a high identity with the known molecules of mammals. The expression level of miR-222 in the testis of Xiang pigs were increased gradually from 1 to 4 months ages. Up to 4 months old age,the expression level was reached its peak value and then decreased. However,the expression pattern of miR-449 b showed an opposite tendency with much low level from 1 to 3 month ages and up to the peak at the fourth month of age. About 420 target genes of miRNA-449 b were predicted,which enriched into 100 KEGG pathways（ P〈0. 05）,such as the oocyte meiosis pathway,Notch signal pathway and Ras signaling pathway.A total of 429 genes were controlled by miRNA-221 involved in 69 KEGG pathways（ P〈 0. 05）,including the cell cycle,Fox O signaling pathways and TGF-beta signal pathway. The miR-449 b and miR-222 target genes were also shared 42 pathways together,which include Wnt signaling pathway and MAPK signaling pathway（ P〈0. 05）.