miR-150+/c-Myb-调控PRDM1/Blimp1诱导外周B淋巴瘤细胞向末端B细胞分化机制

结题报告

项目介绍

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基本信息

批准号：
81272634
项目类别：
面上项目
资助金额：
16.0万
负责人：
赵彤
依托单位：
南方医科大学
学科分类：
H0810.淋巴瘤与淋巴细胞疾病
结题年份：
2013
批准年份：
2012
项目状态：
已结题
起止时间：
2013-01-01 至2013-12-31

项目参与者：
陈少红；韩西群；吴自勍；周新华；汤红平；代新珍；钟琳；葛娟；黎相照；
关键词：
诱导分化 miR-150 c-Myb B细胞淋巴瘤 PRDM1

项目摘要

According to the fact that lymphoma is one of the best model exploring induced differentiation, and in mice miR-150 controls transition from pro-B to pre-B in the B lymphocytes development by targeting c-Myb. we detected that the miR-150 was lowly expressed or absent in 8 human lymphomas cell lines compared with normal B lymphocytes. To explore the mechanism by which miR-150 functions in the pathogenesis of human B-cell lymphoma, we transduced lentivirus vector expressing pre-microRNA-150 into cell lines. the preliminary results showed that the expression of BCL6 was downregulated, and PRDM1 was upregulated in the levels of mRNA and protein in the cell lines of Raji and Daudi transduced with miR-150. Immunophenotypically, both cell lines lost the expression of CD10, and Daudi cells re-expressing miR-150 acquired the expression of CD138, the marker of plasma, compared with control. Concurrent, the transcription factor c-Myb, a potential target of miR150, was downregulated in the set of re-expressing miR-150. Next, we will further conduct tests in the different cell lines and in the models of animal in vivo, employ various biological techniques to elucidate underlying the role of miR150 and the associated genes, such as c-Myb and PRDM1, playing in the differentiation therapy of human B-cell lymphomas. We predict that miR-150 will be a new therapeutic target of B-cell lymphoma in the differentiation therapy.

鉴于B淋巴瘤是探究诱导分化最好模式肿瘤，鼠miR-150负性调控c-Myb表达控制B细胞早期分化。本课题组检测了人八株淋巴瘤细胞miR-150表达，证实B淋巴瘤细胞低表达，构建了pre-micRNA-150慢病毒表达载体及miR-150的Raji150+和Daudi150+细胞亚系，发现miR-150＋瘤细胞c-Myb表达减弱，生发中心标记CD10、Bcl6表达降低，浆细胞分化开关基因PRDM1表达明显升高，出现浆细胞标记CD138表达。在以往基础上，本项目进行基因转染和干扰、miRNA转染和表达阻断、体内和体外等实验，调控B淋巴瘤细胞miR-150表达，观测靶基因c-Myb和末端B细胞分化密切相关基因PRDM1等变化，探究miR-150诱导B淋巴瘤细胞再分化机制，用基因表达谱和免疫沉淀及双荧光素酶分析相关信号通路。获得成功，将会为B淋巴瘤诱导分化治疗提供新的分子靶点及新型研究平台。

结项摘要

鉴于B淋巴瘤是探究诱导分化最好模式肿瘤，鼠miR-150负性调控c-Myb表达控制B细胞早期分化。本课题组检测了人八株淋巴瘤细胞miR-150表达，证实B淋巴瘤细胞低表达，构建了pre-micRNA-150慢病毒表达载体及miR-150的Raji150+、Daudi150+和OCI-Ly10150+细胞亚系，发现miR-150＋瘤细胞c-Myb表达减弱，生发中心标记CD10、Bcl6表达降低，浆细胞分化开关基因PRDM1表达明显升高，出现浆细胞标记CD138表达。在以往基础上，本项目进行基因转染和干扰、miRNA转染和表达阻断等实验，调控B淋巴瘤细胞miR-150表达，观测靶基因c-Myb和末端B细胞分化密切相关基因PRDM1等变化，探究miR-150诱导B淋巴瘤细胞再分化机制，用生物信息学和免疫沉淀分析相关信号通路。获得成功，将会为B淋巴瘤诱导分化治疗提供新的分子靶点及新型研究平台。