Survivin表达对鼠髓样树突状细胞成熟的影响

Effect of survivin expression on the maturation of mouse marrow dendritic cells

目的:探讨survivin的表达变化对小鼠骨髓来源树突状细胞分化成熟的影响。方法:无菌条件下取小鼠的骨髓细胞作为树突状前体细胞,用粒细胞巨噬细胞集落刺激因子(Granulocyte macrophage colony-stimulating factor,GM-CSF)和白细胞介素4(Interleukin 4,IL-4)的培养基定向诱导树突状前体细胞分化成未成熟树突状细胞(Immatureden driticcells,iDC),用GM-CSF,IL-4及肿瘤坏死因子-α(Tumor necrosis factor-alpha,TNF-α)定向诱导树突状前体细胞分化成成熟树突状细胞(Matureden driticcells,mDC):运用半定量RT-PCR,Westernblot比较树突状前体细胞、iDC与mDC中survivinm RNA、survivin蛋白的含量变化。结果:RT-PCR检测结果显示,树突状前体细胞survivinmRNA表达值为(0.735±0.081),iDC为(0.596±0.073,P<0.05),mDC为(0.393±0.052,P<0.01);Westernblot检测结果显示:树突状前体细胞survivin蛋白的表达值为(0.767±0.045),iDC的表达值为(0.396±0.016,P<0.05),mDC的表达值为(0.168±0.009,P<0.01)。结论:survivin在树突状前体细胞中高表达,在iDC与mDC中表达量略低;且survivin在iDC中的表达量高于mDC,这个结论为我们的后续实验提供了理论基础。

Objective：To study the effect of survivin expression on the mature process of dendritic cells from mouse bone marrow. Methods：The cells in marrow as the precurosor of the dendritic cells were taken from mouse in bioclean condition. Then the iDC were induced by the precurosor of DCs with GM-CSF and IL-4 in medium. ThemDC were induced by the precurosor of DCs with GM-CSF,IL-4,and TNF-α in medium. Survivin was compared by RT-PCR,and Western-bolt among these three stages. Results：The RT-PCR showed that the expression ratios of survivin mRNA among the precurosor of DCs,iDC,and mDC were（0.735±0.081）,（0.596±0.073,P〈0.05）,and（0.393±0.052, P〈0.01）. Respectively, the Westernblot showed that the expression ratios of survivin proteinum among the above-mentioned DC’s three stages were（0.767±0.045）,（0.396±0.016, P〈0.05）,and（0.168±0.009,P〈0.01）. Conclusion： Compared with thesurvivin in iDC and mDC, survivin in the precurosor of DCs was expressed more highly;And it was more highly expressed in iDC than in mDC. This conclusion is providing fundamental basis for our follow-up experiments.