Study On Deer Antler Top Tissues IGF-1cDNA Cloning,Expression And The Function Of MiR-1in Antler Cell Proliferation

HQ890468.1. The IGF-1R nucleotide homology of Cervus Nippon, Cattle, Caprine, Homo sapiens, Mus was98.0%,94.2%,92.0%,88.0%. Sended the sequence of IGF-1R to GenBank and gained the number JN009669. The fluorescent quantitation PCR results showed that the content of IGF-1and IGF-1R mRNA in deer antler top tissues from high to low is cartilage, epidermis, dermis, mesenchyme. The content of recombinant protein is more than50%. Western blotting analysis showed that amino acid sequence is correct. The analysis of MTT and cell cycle showed that the activity of recombinant protein is significantly.Find out differential expression miR-1from the miRNA chip and transfect it to deer antler cartilage cell. fluorescent quantitation PCR showed that content of miR-1is325.8times and105.5times higher than control group in24h and48h. Transfection is succession. Western blotting analysis showed that IGF-1protein is lower than control group. MTT and cell cycle showed that the growth rate of cell lower than control group. Double luciferase detection showed that miR-1could inhibit IGF-1protein expression in cell.After48h, the inhibited effect is more obvious.It also proves that IGF-1is miR-1target gene.