The Effect Of LncRNA-AK004418 On Myoblast Proliferation And Differentiation

Long non-coding RNA(lnc RNA) is defined as over 200 nucleotides in length with no protein-coding capacity. Based on the previous microarry results of our laboratory, lncRNA AK004418 was screened out due to its high expression in muscle, RNAi and overexpression were carried out to detect the effect of lncRNA-AK004418 on C2C12 myoblasts proliferation and differentiation. The results are as follows:1. The full length cDNA sequence of AK004418 was obtained by RACE technique, AK004418 gene is located at intron 4 of protein-coding gene SYNPO2 and can not encode protein; AK004418 was highly expressed in leg muscle, longissimus muscle and tongue, while SYNPO2 was widely expressed in 13 tissues studied; The expression of AK004418 increased during myoblast differentiation, but the expression of SYNPO2 was the highest at cell differentiated 5 days; AK004418 was located both in nucleus and cytoplasm.2. QRT-PCR, Western Blot, RTCA xCELLigence and immunofluorescence were used to detect the expression of SYNPO2 gene, myoblast proliferation and differentiation after knockdown and overexpression of AK004418. The results showed that AK004418 promoted cell proliferation, inhibited myoblast differentiation and down-regulated the expression of MyHC and MyoG genes, while there is no significant effect on the SYNPO2 gene expression.3. Microarray was used to detect differentially expressed genes after knockdown of AK004418 in C2C12 cells. The results showed that 1357 genes were up-regulated and 1837 genes were down-regulated, the up-regulated differentially expressed geness were mainly involved in myocardial diseases pathway, and down-regulated differentially expressed genes were mainly involved in cell cycle and regulation of actin cytoskeleton pathways. 21 differentially expressed genes were selected and verified by qRT-PCR, and 18 genes were confirmed to be consistent with the microarray results.The results obtained laid a foundation for further research about regulation mechanism of lncRNA AK004418 in myoblast proliferation and differentiation.