The expression of RNA editing deaminase ADAR2 in human glioma cell lines

Abstract

Objective To observe the mRNA expression of ADAR2 in human glioma cell line SHG44,BT325,U251 and in normal human astrocytes(NHA),to observe the effect of phenylacetate (PA) on the expression of ADAR2 in U251 cells.Methods The level of ADAR2 mRNA in glioma cell lines SHG44,BT325,U251 and in normal human astrocytes (NHA) were detected by real-time fluorescent quantitative polymerase chain reaction (PCR) ; the change of ADAR2 mRNA in U251 cells before and after treated by PA were detected by real-time PCR.Use methyl thiazol tetrazolium (MTT) to detect proliferation of U251 cells on which PA works after 24 h,48 h and 72 h.Use Western blotting to detect ADAR2 protein expression before and after PA treated.Results The ADAR2 mRNA expression level in NHA cells was very weak ( 19.9 ± 2.2),in SHG44,BT325 and U251 cells were 35.6 ±2.8,78.8 ±3.2 and 101.3 ±3.5.After PA (3.0 and 5.0mmol/L) treatment for 24 hours,levels of ADAR2 mRNA were 60.3 ± 1.5 and 50.5 ± 1.2 (P ＜ 0.01 ).MTT assays showed that the U251 cell numbers were significantly reduced with increasing PA concentration in a time-and dose-dependent manner (P ＜0.01 ).Western blotting revealed that ADAR2 protein expression was reduced after PA treatment in U251 cells.Conclusion ADAR2 showed different expression levels among different glioma cell lines.PA can inhibit the expression of ADAR2 mRNA and protein in U251 cells. Key words: Glioma; ADAR2; Phenylacetate; Real-time fluorescent quantitative polymerase chain reaction; Western blotting