摘要
目的:探讨大鼠C6胶质瘤细胞中gdnf基因高转录与其启动子Ⅰ区组蛋白乙酰化的关系。方法:应用Real-time PCR和ChIP-PCR技术分别检测了大鼠正常星形胶质细胞和C6胶质瘤细胞中gdnf基因mRNA的表达水平以及其启动子Ⅰ区组蛋白H3K9的乙酰化程度;利用Real-time PCR技术,检测了不同浓度的组蛋白乙酰基转移酶抑制剂姜黄素(Curcumin)或去乙酰化酶抑制剂曲古抑菌素A(TSA)处理对C6胶质瘤细胞中gdnf基因mRNA表达的影响。结果:较之正常星形胶质细胞,C6胶质瘤细胞中gdnf基因mRNA的表达量极显著增高(P<0.01),并且其启动子Ⅰ区H3K9的乙酰化水平也显著升高(P<0.05)。C6胶质瘤细胞经Curcumin处理24 h后,gdnf基因mRNA的表达量随药物浓度的升高而降低,且100μmol/L作用浓度时其表达量下降了74.17%(P<0.001);相反,TSA处理后gdnf基因mRNA的表达量呈上升趋势,且200nmol/L组其表达量约上升145.35%(P<0.05)。结论:在大鼠C6胶质瘤细胞中gdnf基因启动子Ⅰ区H3K9发生了高乙酰化修饰,这种修饰可能是其高转录的原因。
Objective： To investigate the relationship between gdnfmRNA expression and histone acetylation of the promoter re-gion Ⅰ in rat C6 glioma cells. Methods： Real-time PCR and ChlP-PCR techniques were used respectively to detect the expression level of gdnfmRNA and acetylation status of histone H3K9 in the promoter region Ⅰ in normal rat astrocytes and C6 glioma cells; After treating with varying concentrations of histone acetyltransferase inhibitor Curcumin or deacetylase inhibitor trichostatin A （TSA） in C6 glioma cells, the expression ofgdnfmRNA was detected by real-time PCR. Results： Compared with that in the normal astrocytes, the gdnfmRNA expression increased significantly in C6 glioma cells （P〈0.01）, and the acetylation of H3K9 also enhanced in promoter Ⅰ region signifi-cantly （P〈0.05）. At 24 h after treatment with Curcumin, the expression ofgdnfmRNA decreased in concentration-dependent manner in C6 glioma cells, and about 74.17% decreased was observed in 100 μmol/L group （P〈0.001）; In contrary, gdnfmRNA expression rase after TSA treatment, and a near-maximum increase of 145.35 % was observed in 200 nmol/L group （P〈0.05）. Conclusion： H3K9 hyperacety-lation occurred in gdnfpromoter region Ⅰ, which may be the cause of high gdnftranscription in rat C6 glioma cells.
出处
《现代生物医学进展》
CAS
2014年第11期2015-2018,共4页
Progress in Modern Biomedicine
基金
国家自然科学基金项目(31271358)
江苏省自然科学青年基金(SBK201341565)
江苏省博士后基金(1301068C)
枣庄市医药卫生科技发展计划项目(2013005)
枣庄市科学技术发展计划项目(201361)
