摘要
目的以Eμ-BCL10转基因(transgenic,Tg)小鼠为模型研究BCL10高表达引起MALT淋巴瘤发病的分子机制。方法 PCR鉴定Eμ-BCL10转基因小鼠,Western blotting检测BCL10转基因蛋白的表达,组织学方法显示BCL10Tg小鼠脾脏MALT淋巴瘤的前体细胞-边缘带(marginalzone,MZ)B细胞扩增,流式细胞仪鉴定脾脏边缘带B细胞并分离出这群B细胞用于抗凋亡机制的研究,使用Annexin V-FITC/PI双染法经流式细胞仪检测边缘带B细胞凋亡情况。结果 PCR鉴定BCL10转基因阳性(Tg/+)小鼠,Western blotting证明内源性BCL10蛋白在Tg/+和非转基因野生型(widetype,WT)小鼠中表达的水平相同;BCL10Tg/+小鼠BCL10蛋白的表达量是WT小鼠的2倍。组织学染色显示,BCL10Tg/+小鼠脾脏增生的B细胞可能是边缘带B细胞,将这群B细胞用CD21、CD23染色并经流式细胞仪分析,证明这群增生的B细胞正是CD21high/CD23low的边缘带B细胞。为了进一步研究边缘带B细胞的增生机制,采用CD43、CD23双染,然后通过流式细胞仪分选出BCL10Tg/+以及WT小鼠的边缘带B细胞。BCL10Tg/+小鼠的边缘带B细胞群约30%可以在含5%FBS的RPMI-1640中存活1周以上,而WT小鼠的边缘带B细胞群在第3天几乎全部死亡(P<0.01),这一结果说明,BCL10的高表达引起边缘带B细胞的抗凋亡效应。Anti-IgM诱导边缘带B细胞凋亡实验结果显示,BCL10高表达保护anti-IgM诱导的细胞凋亡。结论 BCL10高表达引起脾脏边缘带B细胞扩增,扩增的B细胞具有抗凋亡特性,这种抗凋亡特性影响抗原受体信号传导途径。这些结果可以在一定程度上解释因BCL10高表达而引起的MALT淋巴瘤的发病分子机制。
Objective To explore the molecular mechanisms of MALT lymphoma caused by BCL10 over-expression. Methods BCL10 transgenic （Tg ） mice were identified by BCL10 transgene specific PCR,and BCL10 protein expression were verified by Western Blot. MALT lymphoma precursor cells-marginal zone（MZ） B cells expansion in BCL10 Tg mice were defined by histology methods and flow cytometry analysis,anti-apoptotic effects of separated MZ B cells were examined with Annexin V-FITC / PI staining. Results The BCL10 Tg/＋ mice used in the experiment were verified by transgene specific PCR. Protein expression of transgene is almost at the same level as endogenous BCL10,which means,in Tg/＋ mice,BCL10 protein has doubled than that in non-transgenic WT mice. By histology,we found B cells expansion in Tg/＋ mice which look like MZ B cells morphologically. By verification with CD21,CD23 staining followed by flow cytometry analysis,it is proven those expanded B cells are CD21high/CD23low MZ B cells. To investigate the mechanisms causing the MZ B cells expansion,we separate the MZ B cells from both Tg/＋ and WT control mice by cell sorting with CD43,CD23 double staining. About 30% separated MZ B cells from Tg/＋ mice can survive at least up to one week in RPMI-1640 containing 5% FBS,but those from WT mice almost all died at day 3（P0.01）,this result means over-expression of BCL10 in MZ B cells causes anti-apoptotic effects to those cells. We studied pathways of anti-apoptotic that BCL10 might be involved by inducing apoptosis with anti-IgM,BCL10 over-expression protect anti-IgM caused cell apoptosis. Conclusion Over-expression of BCL10 causes expansion of MZ B cells which have anti-apoptotic property. This property is involved antigen receptor pathway. These results may partially answer the molecular mechanisms of MALT lymphoma with t（1; 14） （p22; q32） chromosome translocation.
出处
《首都医科大学学报》
CAS
北大核心
2010年第3期353-358,共6页
Journal of Capital Medical University
基金
国家自然科学基金(30872933)
北京市自然科学基金(7092011)
北京市教育委员会科技发展计划项目(KM200910025004)资助项目~~
作者简介
Corresponding author, E-mail： quangeng_zhang@ yahoo. com. cn
