BCL10高表达引起脾脏边缘带B细胞抗凋亡而致其体内增生

首都医科大学学报 ›› 2010, Vol. 31 ›› Issue (3): 353-358.

BCL10高表达引起脾脏边缘带B细胞抗凋亡而致其体内增生

武丽峰, 杨伊姝, 陈彦, 闫卓红, 王爽力, 孙闽, 张权庚^*

首都医科大学基础医学院免疫学系

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-06-21 发布日期:2010-06-21
通讯作者: 张权庚

Overexpression of BCL10 is Anti-apoptotic to Splenic Marginal Zone B Cells Resulting in Their Hyperplasia

WU Li-feng, YANG Yi-shu, CHEN Yan, YAN Zhuo-hong, WANG Shuang-li, SUN Min, ZHANG Quan-geng^*

Department of Immunology, School of Basic Medical Sciences Capital Medical University

Received:1900-01-01 Revised:1900-01-01 Online:2010-06-21 Published:2010-06-21
Contact: ZHANG Quan-geng

摘要/Abstract

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BCL10 Tg小鼠脾脏MALT淋巴瘤的前体细胞-边缘带（marginal zone，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（wide type，WT）小鼠中表达的水平相同；BCL10 Tg/+小鼠BCL10蛋白的表达量是WT小鼠的2倍。组织学染色显示，BCL10 Tg/+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BCL10 Tg/+以及WT小鼠的边缘带B细胞。BCL10 Tg/+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。

关键词: BCL10转基因小鼠, MALT淋巴瘤, 边缘带B细胞, 抗凋亡

Abstract:

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（P＜0.01）, 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.

Key words: BCL10 transgenic mice, MALT lymphoma, marginal zone B cell, anti apoptosis

中图分类号:

R 733.7

武丽峰;杨伊姝;陈彦;闫卓红;王爽力;孙闽;张权庚. BCL10高表达引起脾脏边缘带B细胞抗凋亡而致其体内增生[J]. 首都医科大学学报, 2010, 31(3): 353-358.

WU Li-feng;YANG Yi-shu;CHEN Yan;YAN Zhuo-hong;WANG Shuang-li;SUN Min;ZHANG Quan-geng. Overexpression of BCL10 is Anti-apoptotic to Splenic Marginal Zone B Cells Resulting in Their Hyperplasia[J]. Journal of Capital Medical University, 2010, 31(3): 353-358.