ACSS2通过H4K12ac靶向调控SH-SY5Y细胞中mTOR的表达机制

ACSS2 targets the expression mechanism of mTOR in SH-SY5Y cells through H4K12ac

目的 探讨乙酰辅酶A合成酶2(ACSS2)通过组蛋白H4赖氨酸12乙酰化(H4K12ac)靶向调控SH-SY5Y细胞中雷帕霉素靶蛋白(mTOR)的表达机制。方法 构建ACSS2慢病毒载体,感染SH-SY5Y细胞系,建立ACSS2低表达(ACSS2低表达组)、ACSS2过表达(ACSS2过表达组)细胞系和对照细胞系(NC组);采用组蛋白去乙酰化酶抑制剂伏立诺他(SAHA)处理上述细胞系并分别计为ACSS2低表达+SAHA组、ACSS2过表达+SAHA组、NC+SAHA组。采用Western blot法检测各组蛋白相对表达水平。结果 ACSS2过表达组ACSS2、mTOR的蛋白相对表达水平和H4K12ac修饰水平均显著高于ACSS2低表达组和NC组,差异均有统计学意义(t=15.83、26.38、18.56,26.25、19.52、17.21,P均<0.05);且NC组ACSS2、mTOR的蛋白相对表达水平和H4K12ac修饰水平均显著高于ACSS2低表达组,差异均有统计学意义(t=6.83、7.52、13.21,P均<0.05)。转染SAHA后,ACSS2低表达+SAHA组、ACSS2过表达+SAHA组和NC+SAHA组ACSS2表达均出现显著表达变化,两两组间比较差异均有统计学意义(P均<0.05);ACSS2过表达+SAHA组H4K12ac修饰水平和mTOR的蛋白相对表达水平显著高于ACSS2低表达+SAHA组和NC+SAHA组,差异均有统计学意义(t=21.56、11.56,10.21、9.75,P均<0.05),NC+SAHA组H4K12ac修饰水平和mTOR的蛋白相对表达水平高于ACSS2低表达+SAHA组,差异均有统计学意义(t=15.63、12.31,P均<0.05)。结论 ACSS2通过调节H4K12ac修饰水平靶向介导mTOR的表达。

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.