Mechanism Of MiR-10b Targeting PTEN To Promote Apoptosis Of Porcine Ovarian Granulosa Cells

The ovary is an important reproductive organ of mammals,and plays a very important role in ovulation and fertility regulation.This shows that the health of the ovary has a great impact on the reproductive capacity of mammals.The proliferation,apoptosis and significant functional differentiation of ovarian granulosa cells are of great significance to the ovary and are also necessary for follicular maturation.Many studies have shown that miRNA and related genes play an important regulatory role in granule cells function.In the early stage of our research group,through transcriptome and miRNA sequencing analysis of porcine luteal phase and follicular phase ovarian tissue,key miRNAs and m RNAs that may affect the transition from luteal phase to follicular phase were selected,and candidate gene PTEN was selected.And through bioinformatics methods to find miR-10 b that may have a targeting relationship with PTEN.Through the use of Annexin VFITC/PI flow detection,qPCR,Western blot and other technologies,the effect of PTEN on the function of porcine ovarian granulosa cells and its miRNA targeting function were studied,and the dual luciferase reporter gene system was used to verify the PTEN have targeting relationship between miR-10 b.The purpose of this experiment was to clarify the function and mechanism of miR-10 b on the apoptosis of porcine ovarian granulosa cells,in order to gain a deeper understanding of the apoptosis mechanism of porcine ovarian granulosa cells,lay a foundation for elucidating the genetic regulation of reproductive endocrine,and further study follicular atresia.The specific results of this study are as follows:1.The results of Annexin V-FITC/PI flow cytometry showed that miR-10 b and PTEN can promote the apoptosis of porcine ovarian granulosa cells.2.qPCR and Western blot results showed that miR-10 b negatively regulates PTEN expression at the transcriptional and translational levels.3.Using bioinformatics analysis,it was found that miR-10 b is highly conserved among different species,and predict the binding site of miR-10 b to PTEN.The results of using the dual luciferase reporter gene system showed that miR-10 b can target the PTEN 3'UTR region and inhibit the reporter gene fluorescent activity.The above results indicate that there is a targeting relationship between miR-10 b and PTEN,and miR-10 b promotes the apoptosis of porcine ovarian granulosa cells by targeting PTEN.