Screening And Identification Of FGF5 Regulated Genes Related To Villus Growth In Liaoning Cashmere

In China,cashmere is a kind of important import and export commodity.Liaoning cashmere goats are also known for high quality cashmere,cashmere yield and stable genetic properties.It is an unique treasure of China.Although this population is cultivated in the1980 s,little is known about the molecular mechanism and regulatory mechanism of cashmere growth.So the experiment has been studied and discussed.In this study,we first treat with FGF5 at different concentrations and for different periods of time to deal with skin flbroblast from Liaoning Cashmere goats.We use the RNA sequencing to extract and analyze total RNA.And then we select and analyze the differential express genes and the biological processes.Through statistical analysis and bioinformatics analysis,the main results of this experiment are as follows:(1)In this study,we set up four treatment groups.The three experimental groups is:F4＿24H group,F4＿72H group,F6＿24H group,and one control group: C group.Using RNA-seq to analyze it.The results show that 164 differentially expressed transcripts are obtained in the F4＿24h vs C group,with 70 up-regulated transcripts and 94 down-regulated transcripts;F4＿72h vs.C,189 differentially expressed transcripts are obtained,with 78up-regulated transcripts and 111 down-regulated transcripts;F6＿24h vs.C,123 differentially expressed transcripts,27 up-regulated transcripts,and down-regulated transcription There are96.(2)Using GO enrichment analysis,the results show that the differentially expressed genes in the F4＿24H vs C group are mainly enriched in the extracellular matrix in the cell composition and metalloendopeptidase activity,metallo peptidase activity in molecular function..(3)Using the KEGG database for comparison,the results show that the differentially expressed genes are significantly enriched in the three signaling pathways of ECM,FAK and PI3K-Akt in the F4＿24H vs C group,and there are 28 differentially expressed genes,of which TGA6,The OPN,ITGB8,THBS1,ITGA4,ITGA8,LAMA2,and LAMA4 genes are present in the three signaling pathways of ECM,PI3 K,and FAK.According to the genetic information query results on NCBI and GENE CARD website,the expression of ITGA6,OPN,THBS1,and LAMA2 genes and the site of action are related to the growth and development of hair follicles.(4)Analysis of protein network interaction shows that the results of the F4＿72H vs C treatment group have the most significant effect on FGF5 in skin cells of Liaoning Cashmere goats.(5)The expression levels of OPN,ITGAV,ITGA4 and ITGA9 are verified by real-time fluorescence quantitative PCR.The results show significant changes in OPN and ITGAV and were consistent with previous sequencing analyses.(6)Experiments using CCK-8 technique show that the proliferation of FGF5-treated cells is most evident at 1 day(24 h)compared to the control group,and there is no significant change in late proliferation.(7)Analysis of ITGAV and OPN using bioinformatics: OPN is an extracellular protein and ITGAV is a transmembrane protein.