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目的阐明Hes5通过调节哪些蛋白的表达而抑制神经细胞的分化。方法利用蛋白组学同位素标记相对和绝对定量技术(iTRAQ技术)观察Hes5敲除生后8d小鼠室下带蛋白表达的变化,并用Western blotting技术对表达上调和下调的蛋白进行验证。结果在Hes5敲除生后8d小鼠室下带发现19种蛋白有重要的变化。与对照组野生型小鼠相比,其中10种蛋白的表达是上调的,变化的倍率在1.20~1.32之间。9种蛋白的表达是下调的,变化的倍率在0.77~0.83之间。上调的蛋白主要与促进神经细胞的生长,细胞的迁移,黏附和信号的转导有关,而下调的蛋白主要是抑制细胞的生长和维持细胞骨架的蛋白。Western blotting验证蛋白表达的变化与iTRAQ结果一致。结论Hes5可能通过影响与细胞生长、迁移以及细胞骨架等有关的多种蛋白的表达来影响细胞的分化。
Objective To elucidate the role of Hes5 in inhibiting neuronal differentiation by regulating the expression of these proteins. Methods The protein expression of subventricular zone in 8-day-old Hes5 knockout mice was detected by proteomic isotope labeling relative and absolute quantification (iTRAQ) technique. Western blotting was used to verify the up-regulated and down-regulated proteins. RESULTS: Nineteen proteins were found to have important changes in the subventricular zone of mice 8 days after Hes5 knockout. Compared with the wild type mice in the control group, the expression of 10 proteins was up-regulated, with the rate of change ranging from 1.20 to 1.32. The expression of 9 kinds of proteins is down-regulated, the rate of change is between 0.77 ~ 0.83. Up-regulated proteins mainly promote nerve cell growth, cell migration, adhesion and signal transduction, while down-regulated proteins are mainly those that inhibit cell growth and maintain the cytoskeleton. Western blotting confirmed the change in protein expression was consistent with the iTRAQ results. Conclusion Hes5 may influence cell differentiation by affecting the expression of various proteins related to cell growth, migration and cytoskeleton.