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目的:研究Ikaros在调节垂体生长激素(GH)基因表达的作用。方法:应用系列的细胞与分子生物学技术如细胞培养、稳定转染、免疫组化、Northern Blot(N.B)、蛋白质印记、染色质免疫共沉淀等方法分析Ik1及其同工蛋白Ik-6在垂体GH基因表达调节中的作用。结果:野生型Ikaros(Ik1)在垂体GH4细胞中抑制了GH基因mRNA和蛋白的表达水平。Ikaros不直接与GH基因启动子结合,但能抑制TSA引起的GH基因启动子的组蛋白乙酰化作用。Ik1能够选择性地抑制GH启动区的染色质组蛋白-3的乙酰化作用,因而阻碍了Pit1与GH启动子DNA的结合。结论:Ikaros在垂体激素基因表达过程中抑制了GH基因的表达(分泌),这一研究为功能性垂体腺瘤GH过度分泌的分子靶向治疗提供了新的参考。
Objective: To study the role of Ikaros in regulating pituitary growth hormone (GH) gene expression. METHODS: Ik1 and its isoforms Ik-6 were analyzed using a series of cell and molecular biology techniques such as cell culture, stable transfection, immunohistochemistry, Northern Blot (NB), Western blotting, and chromatin immunoprecipitation Role of pituitary GH gene expression regulation. Results: Wild-type Ikaros (Ik1) inhibited the expression of GH mRNA and protein in pituitary GH4 cells. Ikaros does not directly bind to the GH gene promoter, but inhibits the histone acetylation of the GH gene promoter caused by TSA. Ik1 selectively inhibits the acetylation of chromatin Histone-3 in the GH-priming region and thus blocks the binding of Pit1 to GH promoter DNA. CONCLUSION: Ikaros inhibits the expression (secretion) of GH gene in the process of pituitary hormone gene expression. This study provides a new reference for the molecular targeted therapy of GH hypersecretion in functional pituitary adenoma.