免疫球蛋白κJ区重组信号结合蛋白对CD133阳性室管膜细胞增殖与分化的影响

目的 探讨免疫球蛋白κJ区的重组信号结合蛋白(RBP-JK)对CD133阳性室管膜细胞增殖与分化的影响及可能的机制.方法 分离孕12 d的美国癌症研究所(ICR)胚胎小鼠(3只)侧脑室室管膜细胞进行原代培养,并使用 RBP-Jκ-siRNA 干扰 RBP-JK,以及选用2～3月龄体重为20～25 g 的 CD133-C...

Effect of recombination signal binding protein for immunoglobulin Kappa J region on the proliferation and differentiation of CD133-positive ependymal cells

Objective To explore the effect of recombination signal binding protein for immunoglobulin Kappa J region (RBP-Jκ) on the proliferation and differentiation of CD133-positive ependymal cells and its possible mechanism. Methods RBP-Jκ in CD133-positive ependymal cells of lateral ventricle was interfered with siRNA in the fetuses of embryos which were isolated from 12-day pregnant Institute of Cancer Research(ICR) mouse (3 mice) and knocked out in CD133-CreERTM: :ROSA26-LacZ: :RBP-Jκflox/floxmouse(3 mice) treated with tamoxifen (TAM) intraperitoneally. The proliferation and differentiation of CD133-positive ependymal cells, as well as the related molecules in RBP-Jκ upstream or downstream were evaluated and measured by immunofluorescent staining, Real-time PCR and Western blotting. ResultsBy interfering or knocking out RBP-Jκ, the number of CD133 or β-galactosidase(β-GAL)/doublecortin(DCX) β-tubulin Ⅲ or CD133 β-GAL/microtubule associated protein-2 (MAP-2) neuronal nuclear antigen(NeuN) or CD133 β-GAL/proliferating cell nuclear antigen(PCNA) double-positive cells were significantly increased in vitro or in vivo. Meanwhile, the result from Real-time PCR and Western blotting showed that RBP-Jκ and hairy and enhancer of split1(Hes1) mRNA and their protein expression levels decreased significantly, while Notch signaling pathway receptor 1(Notch1) mRNA and its protein expression levels increased significantly. Conclusion The interference or knockout of RBP-Jκ will lead to upregulateing Notch1 and downregulate Hes1 expression levels, which eventually increase the proliferation and differentiation of CD133-positive ependymal cells.