Regulating Mechanism Of MiR-24-3p In Myoblasts Proliferation And Apoptosis

Orizontally continuously increase along with the research in modern molecular biology and cell biology,beef cattle breeding in China has gradually estepped into the stage of emerging biotechnology breeding.Muscle development is one of the most watched segments in beef cattle production,and its research is also gradually deepening,from molecular marker-assisted selection at the DNA level to the study of the key gene function at the mRNA level,and then extended to the study of the regulatory mechanism of non-coding RNA at the epigenetic level.miRNA is a small fragment of non-coding RNA,and its classical mode of action is to regulate related biological functions by targeting binding mRNA 3 '-UTR.In recent years,numerous miRNAs related to muscle development have been found,but there are no reports on the existence of relevant miRNA that conservatively regulates muscle development among different species.Therefore,in this study,based on high-throughput sequencing results of miRNAs in muscle tissues of different species(cattle,rats,goats and pigs),Venn chart found that miR-24-3p may be a potential miRNA that can conservative regulate muscle development.To confirm its biological function in muscle development,C2C12 cell lines and bovine primary muscle cells were tested.In order to explore the conservative regulation mechanism of miR-24-3p,the dual-luciferase reporter system was used to verify the targeting relationship between miR-24-3p and the common target between species,and to clarify the molecular mechanism of miR-24-3p regulating the biological process related to muscle development by extensively regulating the common target gene group between species.The results as follow:1.The miRNA transcriptome sequencing data from muscle tissues of cattle,rats,goats and pigs were downloaded for Venn analysis,and miR-24-3p could be a potential target for conservative regulation of muscle development among species..2.C2C12 cells were used as models to overexpress or inhibit the expression of mmu-miR-24-3p.CCK-8 and qRT-PCR techniques showed that mmu-miR-24-3p could positively regulate the proliferation of C2C12 cells by regulating key proliferation and apoptosis genes in the process of muscle development.3.Bovine muscle primary cell was used as model to overexpress or inhibit the expression of bta-miR-24-3p.CCK-8 and qRT-PCR showed that bta-miR-24-3p could also positively regulate the proliferation of bovine muscle primary cells by regulating key proliferation and apoptosis genes in the process of muscle development.4.Targetsan software predicted the target genes of miR-24-3p in cattle,mouse,humansand rats.Venn analysis found that there were a staggering proportion of common target genes among the four species,and the muscle development related target genes were verified by using dual-luciferase reporter system and qRT-PCR technology,and it was found that miR-24-3p could regulate the 3 '-UTR of most predicted target genes.To sum up,this study is the first to start from the perspective of inter-species conserved regulation of miRNA in muscle development,and found that miR-24-3p has the function of inter-species conservative regulation of muscle development.The molecular mechanism of miR-24-3p was explored,and it was found that the conserved regulation of miR-24-3p on muscle development among species may be achieved through the network regulation of its large-scale shared target genes.This study provides a new idea for further research on the regulation of miRNA on muscle development,and provides a certain theoretical and scientific basis for beef cattle breeding.