ACSS2 targets the expression mechanism of mTOR in SH-SY5Y cells through H4K12ac
Objective To explore the mechanism of Acetyl-coa synthetase 2(ACSS2)targeting and regulating the expression of mammalian target of rapamycin(mTOR)in SH-SY5Y cells through histone H4 lysine 12 acetylation(H4K12ac).Methods Construct ACSS2 lentiviral vector,infect SH-SY5Y cell line,establish ACSS2 underexpression(ACSS2 underexpression group),ACSS2 overexpression group(ACSS2 overexpression group)and control cell lines(NC group).The above cell lines were treated with the histone deacetylase inhibitor volinotha(SAHA)and classified into ACSS2 underexpression+SAHA group,ACSS2 overexpression+SAHA group and NC+SAHA group,respectively.The relative expression levels of ACSS2 and mTOR protein and the modification levels of H4K12ac in each group were detected by Western blot.Results The relative expression level of ACSS2 and mTOR protein and the modification level of H4K12ac in ACSS2 overexpression group were significantly higher than those in ACSS2 underexpression group and NC group,the differences were statistically significant(t=15.83,26.38,18.56;26.25,19.52,17.21;all P＜0.05);and the relative expression level of ACSS2 and mTOR protein and the modification level of H4K12ac in NC group were significantly higher than the ACSS2 underexpression group,the differences were statistically significant(t=6.83,7.52,13.21;all P＜0.05).After transfection with SAHA,the expression of ACSS2 in ACSS2 underexpression+SAHA group,ACSS2 overexpression+SAHA group and NC+SAHA group showed significant changes,the differences between the two groups were statistically significant(all P＜0.05);The H4K12ac modification level and the relative expression level of mTOR protein in ACSS2 overexpression+SAHA group were significantly higher than those in ACSS2 underexpression+SAHA group and NC+SAHA group,the differences were statistically significant(t=21.56,11.56;10.21,9.75;all P＜0.05);The H4K12ac modification level and the relative expression level of mTOR protein in NC+SAHA group were significantly higher than those in ACSS2 underexpression+SAHA group,the difference was statistically significant(t=15.63,12.31;both P＜0.05).Conclusion ACSS2 might target the expression of mTOR by regulating the level of H4K12ac modification.
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