miR-27b介导Pax3基因调控山羊骨骼肌细胞增殖与分化研究

肌肉生长发育表现为肌纤维数目增加和细胞体积膨胀，这两个方面依赖于肌肉细胞的增殖和分化。而调控肌肉细胞增殖与分化分子机制，尤其是转录后调节机制尚不明确。microRNAs的发现为此研究提出新的方向。本组前期研究发现在山羊不同发育阶段骨骼肌中miR-27b显著差异表达，结合最新资料推测miR-27b通过作用Pax3基因调控骨骼肌细胞的增殖与分化。本项目拟以前期筛选的miR-27b为基础，以生长较慢的黄淮山羊和生长快的波尔山羊为材料，分析miR-27b及其预测靶基因Pax3在山羊骨骼肌及其细胞中的表达规律；进而通过双荧光素酶活性实验、qPCR、Western blot等方法研究miR-27b靶向作用Pax3基因调控山羊骨骼肌细胞增殖与分化作用及机制。该项研究在理论上可望在肌肉细胞定向分化分子机制等方面取得突破性发现，为进一步揭示骨骼肌生长发育相关基因调控研究提供新的探索途径和理论基础。

Muscle growth and development were due to an increase in the number of muscle fibers and cell volume expansion which dependent on the proliferation and differentiation of muscle cells. However, the regulation mechanisms, especially in post-transcriptional level, were unclear. The discovery of microRNAs provided a new direction on the study. Our previous studies found significant differences in expression of miR-27b in skeletal muscle of goats at different developmental stages. Combined with other research, we proposed that miR-27b regulated the skeletal muscle cell proliferation and differentiation through Pax3 gene. To further investigate the regulation mechnism, we will analyze the expression pattern of miR-27b and Pax3 in both muscle samples and skeletal muscle cells of Huanghuai Goat. Using the dual luciferase activity assays, Realtime Q-PCR, Western blot and confocal microscopy observation, we will study the molecular mechanisms of goat skeletal muscle cell proliferation and differentiation modified by miR-27b regulation of Pax3 expression. The study is expected to have some breakthrough discovery about directional differentiation mechanism of the skeletal muscle cells. The results will provide new avenues of exploration for further reveal gene regulation in the skeletal muscle growth and development related to studies.