组蛋白乙酰化修饰对大鼠C6胶质瘤细胞中GDNF基因转录调控的研究

发布时间：2018-03-05 14:39

本文选题：胶质瘤　切入点：GDNF　出处：《苏州大学》2014年硕士论文　论文类型：学位论文

【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区组蛋白乙酰化修饰的关系。方法：应用RT-PCR、Real-time PCR和ChIP-PCR技术分别检测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区和Ⅱ区组蛋白H3K9的乙酰化程度；利用Real-time PCR和ChIP-PCR技术，检测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表达以及其启动子Ⅰ区和Ⅱ区组蛋白H3K9的乙酰化的影响。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（p0.01），并且其启动子Ⅰ区和Ⅱ区组蛋白H3K9的乙酰化水平也显著升高（P0.05）。C6胶质瘤细胞经Curcumin处理24h后，GDNF基因mRNA的表达量随药物浓度的升高而降低，且100μM作用浓度时其表达量下降了74.17%（P0.001）；相反，，TSA处理后GDNF基因mRNA的表达量呈上升趋势，且200nM组其表达量约上升145.35%（P0.05）。同时，GDNF基因启动子Ⅱ区组蛋白H3K9的乙酰化修饰水平对Curcumin或TSA的作用敏感，其乙酰化的变化趋势与转录水平变化趋势一致。结论：在胶质瘤细胞中，GDNF基因启动子Ⅰ区和Ⅱ区组蛋白H3K9乙酰化水平显著增高；GDNF基因启动子Ⅱ区组蛋白H3K9高乙酰化可能是该基因异常高转录的原因之一。
[Abstract]:Objective: To investigate the relationship between the high transcription of GDNF gene in rat C6 glioma cells and the modification of histone acetylation in the promoter region.
Methods: RT-PCR was used to detect the expression level of GDNF in normal rat astrocytes and C6 glioma cells in mRNA gene and its promoter region I and II histone H3K9 acetylation level of Real-time PCR and ChIP-PCR technology; using Real-time PCR and ChIP-PCR technology, detection of histone acetyl transferase in different concentration curcumin (Curcumin) inhibitors or histone deacetylase inhibitor trichostatin A (TSA) on the expression of GDNF C6 gene in mRNA glioma cells as well as the influence of the promoter region I and II histone H3K9 acetylation.
Results: compared with normal astrocytes. The expression of GDNF gene mRNA in C6 glioma cells was significantly increased (P0.01), the promoter region I and II histone H3K9 acetylation level also significantly increased (P0.05) and.C6 glioma cells treated by Curcumin 24h, and GDNF gene expression in mRNA decreases with the increase of drug concentration, 100 M concentration and its expression decreased by 74.17% (P0.001); on the contrary, after the treatment of TSA and GDNF gene expression of mRNA increased and the expression level of 200nM group increased by approximately 145.35% (P0.05). At the same time, the GDNF gene promoter function level of acetylation sub District II histone H3K9 on Curcumin or TSA sensitive, consistent variation trend and trend of the transcription level of acetylation.
Conclusion: in the glioma cells, the acetylation level of histone H3K9 in GDNF gene promoter region and II region increased significantly, and the histone H3K9 high acetylation in GDNF gene promoter region might be one of the reasons for the abnormal high transcription of the gene.

【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R739.4

【共引文献】