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作 者:黎衍敏[1] 苟欣[1] 何卫阳[1] 王海峰[1]
机构地区:[1]重庆医科大学附属第一医院泌尿外科,重庆400016
出 处:《重庆医科大学学报》2009年第9期1144-1147,共4页Journal of Chongqing Medical University
基 金:国家自然科学基金资助项目(30772043)
摘 要: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.
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