Modulatory effects of acetylation of histone H3 on Nanog gene in glioma

Li Zhongying; Niu Chaoshi; Wang Yan; Yang Yang; He Hu; Li Dongxue

doi:10.11850/j.issn.1009-122X.2014.07.013

Abstract：Objective To investigate the modulatory effects of acetylation of histone H3 on the proliferation-related factor Nanog in glioma. Methods U87 glioma cells were cultured in vitro and then exposed to apicidin at different concentrations and different time points as experimental group, while U87 cells exposed to the dimethylsulfoxide (DMSO) as DMSO group and not receiving any solvent as blank control group. MTT assay was used to detect the effect of apicidin on cell proliferation and select suitable intervention concentration. Real-time PCR was employed to measure the Nanog mRNA expression. Western blot was applied to detect the acetylation of histone H3. Chromatin immunoprecipitation real-time PCR was used to measure the level of the acetylation of histone H3 in promoter region of Nanog. Results Apicidin significantly inhibited the proliferation of U87 cells by MTT assay, and the half maximal inhibitory concentration (IC50) was 1.74±0.13 μmol/L in the experimental group. Compared with the blank control group, the Nanog mRNA expression of U87 glioma cells decreased by 46.52%±0.53% in the experimental group (P<0.05), the expressions of acetylation of histone H3 and Nanog protein in U87 glioma cells also decreased obviously (P<0.05), and the acetylation of histone H3 expression in the promoter region of Nanog decreased by 46.52%±0.82%(P<0.05). Conclusions Low level of the acetylation of histone H3 can inhibit Nanog expression, and Nanog expression is regulated by the acetylation of histone H3 in glioma cell line.