Jian Xue1, Jingwen Xue2, Ji Zhang3, Dan Li4, Lei Jiang4. 1. College of Animal Husbandry & Veterinary, Jinzhou Medical University, No. 48, Section 5, Renmin Street, Guta District, Jinzhou City, 121001, Liaoning Province, People's Republic of China. jianx@163.com. 2. College of Animal Science and Veterinary Medicine, Shandong Agricultural University, Tai'an, 271018, People's Republic of China. 3. College of Food Science and Engineering, Jinzhou Medical University, Jinzhou City, 121001, People's Republic of China. 4. College of Animal Husbandry & Veterinary, Jinzhou Medical University, No. 48, Section 5, Renmin Street, Guta District, Jinzhou City, 121001, Liaoning Province, People's Republic of China.
Abstract
OBJECTIVES: To explore the roles of miR-130b-3p and miR-301b-3p which may regulate Rb1-inducible coiled-coil 1 (Rb1cc1) expression during myogenic differentiation of chicken primary myoblasts. RESULTS: After 4 days of myogenic differentiation, myotubes appeared and after 6 days the cells fused to each other and expression of MyHC could be detected by immunofluorescence staining. TargetScan and RNAhybrid 2.2 showed miR-130b-3p and miR-301b-3p were well complementary with the target site of Rb1cc1 3'-untranslated region (3'-UTR). Using the dual-luciferase assay, we found miR-130b-3p and miR-301b-3p could inhibit Rb1cc1 expression by binding to its 3'-UTR. Real-time PCR showed Rb1cc1 mRNA expression level was almost reciprocal to that of miR-130b-3p or miR-301b-3p during myogenic differentiation. Furthermore, over-expression of miR-130b-3p or miR-301b-3p down-regulated the expression levels of Rb1cc1, myoblast determination protein, myogenin and myosin heavy chain. CONCLUSIONS: miR-130b-3p or miR-301b-3p negatively regulate Rb1cc1 expression to affect myogenic differentiation.
OBJECTIVES: To explore the roles of miR-130b-3p and miR-301b-3p which may regulate Rb1-inducible coiled-coil 1 (Rb1cc1) expression during myogenic differentiation of chicken primary myoblasts. RESULTS: After 4 days of myogenic differentiation, myotubes appeared and after 6 days the cells fused to each other and expression of MyHC could be detected by immunofluorescence staining. TargetScan and RNAhybrid 2.2 showed miR-130b-3p and miR-301b-3p were well complementary with the target site of Rb1cc1 3'-untranslated region (3'-UTR). Using the dual-luciferase assay, we found miR-130b-3p and miR-301b-3p could inhibit Rb1cc1 expression by binding to its 3'-UTR. Real-time PCR showed Rb1cc1 mRNA expression level was almost reciprocal to that of miR-130b-3p or miR-301b-3p during myogenic differentiation. Furthermore, over-expression of miR-130b-3p or miR-301b-3p down-regulated the expression levels of Rb1cc1, myoblast determination protein, myogenin and myosin heavy chain. CONCLUSIONS:miR-130b-3p or miR-301b-3p negatively regulate Rb1cc1 expression to affect myogenic differentiation.