期刊论文

Knock-down of RACK1 Inhibits the Expressions of Myog and MHC Genes in C2C12 Myoblast

中文

中国畜牧杂志

0258-7033

2017

53

3

106-111

10.19556/j.0258-7033.2017-03-106

湖北省自然科学资助项目（2015CFB514） 湖北省高等学校优秀中青年科技创新团队计划项目（T201508）

本校为第一机构

本实验旨在研究RNA干扰（RNAi）RACK1基因对C2C12成肌细胞中肌分化标志基因MHC和MyoG表达的影响。将人工合成的靶向RAKC1基因的3条si RNA（1,2,3）转染C2C12细胞,用RT-qPCR方法检测并筛选干扰效率最高的1条siRNA。将筛选出的si RNA转染C2C12成肌细胞并诱导细胞分化,通过RT-qPCR、Western blot和免疫荧光方法在mRNA及蛋白水平检测RACK1基因沉默后对MHC和MyoG表达的影响。此外,Western blot方法检测RACK1基因沉默后对PI3K/Akt和Erk/MAPK通路激活的影响。结果表明：RACK1-si RNA-2干扰效率最高,转染48 h后抑制率可达70%。随后,C2C12细胞转染si RNA-2,诱导分化48h后RTqPCR表明,MHC和MyoG的m RNA表达...

The purpose of the study was to investigate the influence of RACK1 genesilencing by RNA interference（RNAi）on the expressions of two myogenic differentiation markgenes（MHC and Myo G） in C2C12 myoblast. We transfected three chemica lly synthesized RACK1 si RNA（si RNA-1,2,3） into C2C12 myoblasts, and detected the interference efficiency with RT-q PCR in order toselect thesi RNA with the highest interference efficiency. Then, we transfected the selected si RNA into C2C12 cells and induced C2C12 differentiation. In order to study the effect of...