小鼠胸腺雄激素受体的定位及其与新分子Rwdd1的关系

目的 研究雄激素受体(AR)在胸腺萎缩中的作用机制.方法 用免疫组化和RT-PCR观察AR在小鼠胸腺内的分布;用刀豆蛋白A(ConA)诱导胸腺细胞增殖,观察睾酮对细胞增殖的影响;构建AR和Rwdd1的真核表达载体pcDNA3.1-AR和pcDNA3.1-Rwdd1-V5,共转染293T细胞,通过免疫共沉淀(CoIP)法观察二者之间在细胞内是否存在相互作用.结果 AR广泛分布在胸腺皮质和髓质,且主要定位于胞核.在不同胸腺细胞亚群中均有AR表达,其中以CD4-CD8-(DN)细胞表达水平最高.3种胸腺上皮细胞系中仅皮质来源的427.1细胞系存在AR表达.胸腺细胞中存在功能性AR.AR与Rwdd1之间不存在直接相互作用.结论 小鼠胸腺细胞中存在AR的表达,目前尚不能证明AR与Rwdd1存在直接的相互作用.

Distribution of androgen receptor in murine thymus and its relationship with a novel protein Rwdd1

Objective To study the potential role of AR in T cell development.Methods AR expression in thymic cells was detected by immunohistological techniques and RT-PCR amplification.Function of AR was further evaluated by ConA-induced thymocytes proliferation following treatment of increasing dose of testosterone.Eucaryotic expression vectors pcDNA3.1-AR and pcDNA3.1-Rwdd1-V5 were constructed and used to co-transfect 293T cells,and coimmunoprecipitation(CoIP) was carried out to explore the potential interaction between AR and Rwdd1.Results There was AR distribution in both the cortex and medulla part of thymus,and AR located primarily in the nucli of thymic cells.RT-PCR showed that CD4-CD8-(DN) cells had the highest AR expression level among different thymocyte subsets.Of the three thymic epithelial cell lines,only 427.1 which originated from the cortex had AR expression.The existence of functional AR was further confirmed by in vitro ConA stimulated thymocytes proliferation test under testosterone treatment.However,potential interaction between AR and Rwdd1 was not proved by CoIP test.Conclusion There is AR expression in murine thymic cells.No evidence of direct interaction between AR and Rwdd1 is shown yet.