Identification And Functional Study Of LncRNA Related To Growth And Development Of Porcine Skeletal Muscle

Pork is one of the most important meat sources in the world,and its meat quality has an important impact on human nutrition and health.The growth and development of porcine skeletal muscle is one of the important factors affecting meat quality.Therefore,in-depth research on the growth and development mechanism of porcine skeletal muscle has important scientific significance and application value.Long strand non coding RNA(lncRNA)is a class of RNA molecules with a length greater than 200 nt,which plays an important role in cell regulation,including post transcriptional regulation,epigenetic regulation and signal pathway regulation.In organisms,lncRNA plays an important role in cell proliferation,differentiation,migration,and other life processes.Therefore,the identification and functional research of lncRNA related to growth and development of porcine skeletal muscle is of great significance for revealing its mechanism to regulate growth and development of porcine skeletal muscle.In the present study,muscle specific differential expressed lncRNAs were screened through transcriptome sequencing and their functions were also verified.The results from the present research provided a theoretical basis for studying the growth and development mechanism of porcine skeletal muscle.The main results of this study are as follows:1.Transcriptome sequencing was performed on different tissue samples from four different breeds of pigs at different days of age.Two lncRNAs with skeletal muscle specific expression,lncRNA-30095 and lncRNA-3389,were found,and the differential expression of lncRNA-30095 and lncRNA-3389 in skeletal muscle between Bama xiang pigs and Duroc pigs were also verified at the transcriptome level.2.The overexpression and interference vector of lncRNA-30095 was constructed and transfected into porcine skeletal muscle cells.The changes of cell proliferation(PCNA,Ki67,Cyclin A,Cyclin D2),differentiation(Myo G,My HC),and migration marker genes(FHL1,ACTG1)were detected by q PCR and WB.At the same time,the impacts of lncRNA-30095 on porcine skeletal muscle cell proliferation,differentiation and migration were determined by using CCK-8 and Ed U kits,immunofluorescence and Transwell methods,respectively.The results showed that lncRNA-30095 could promote the proliferation of porcine skeletal muscle cells and inhibit their differentiation and migration.3.The si RNA of lncRNA-3389 was constructed and transfected into porcine skeletal muscle cells to detect its effects on the cell proliferation,differentiation,and migration of porcine skeletal muscle.In addition,the interaction between lncRNA-3389 and mi R-133a-3p was also explored.The results showed that lncRNA-3389 can recruit mi R-133a-3p to jointly promote the proliferation of porcine skeletal muscle cells,and inhibit their differentiation and migration.In summary,two skeletal muscle specific expressed lncRNAs,lncRNA-30095 and lncRNA-3389 were screened and their differential expression in Bama and Duroc pigs was confirmed in present study.It was verified that lncRNA-30095 and lncRNA-3389 could regulate the cell proliferation,differentiation and migration of porcine skeletal muscle,and further found that the interaction between lncRNA-3389 and mi R-133a-3p can promote the development of porcine skeletal muscle cells.