Zhang Chen-Guang, Wang Hui, Niu Zhi-Guo, Zhang Jing-Jing, Yin Ming-Mei, Gao Zhi-Tao, Hu Li-Hua
Clinical Laboratory Medicine Department, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei, China.
Asian Pac J Cancer Prev. 2013;14(1):359-65. doi: 10.7314/apjcp.2013.14.1.359.
The high mobility group box 1 (HMGB1) protein is a multifunctional cytokine-like molecule that plays an important role in the pathogenesis of tumors. In this study, real-time polymerase chain reactions and Western blot assays indicated that HMGB1 transcriptional activity and protein level are increased in Tax+-T cells (TaxP). To clarify the mechanisms, a series of HMGB1 deletion reporter plasmids (pHLuc1 to pHLuc6) were transfected into Tax--T cells (TaxN, Jurkat) and Tax+-T cells (TaxP). We found that promoter activity in Tax+-T cells to be higher than that in Tax--T cells, indicating a significant increase in pHLuc6. Bay11-7082 (NF-κB inhibitor) treatment did not block the enhancing effect. Chromatin immunoprecipitation assays revealed that Tax was retained on a HMGB1 promoter fragment encompassing -1163 to -975. Bioinformatics analysis showed six characteristic cis-elements for CdxA, AP-1, AML-1a, USF, v-Myb, and C/EBP in the fragment in question. Mutation of cis- elements for C/EBP reduced significant HMGB1 promoter activity induced by Tax. These findings indicate that Tax enhances the expression of HMGB1 gene at the transcriptional level, possibly by interacting with C/EBP.
高迁移率族蛋白B1(HMGB1)是一种多功能细胞因子样分子,在肿瘤发病机制中起重要作用。在本研究中,实时聚合酶链反应和蛋白质印迹分析表明,在Tax阳性T细胞(TaxP)中,HMGB1的转录活性和蛋白水平升高。为阐明其机制,将一系列HMGB1缺失报告质粒(pHLuc1至pHLuc6)转染至Tax阴性T细胞(TaxN,Jurkat细胞)和Tax阳性T细胞(TaxP)中。我们发现Tax阳性T细胞中的启动子活性高于Tax阴性T细胞,表明pHLuc6显著增加。Bay11 - 7082(NF-κB抑制剂)处理并未阻断这种增强作用。染色质免疫沉淀分析显示,Tax保留在包含-1163至-975的HMGB1启动子片段上。生物信息学分析表明,该片段中存在针对CdxA、AP-1、AML-1a、USF、v-Myb和C/EBP的六个特征性顺式元件。C/EBP顺式元件的突变降低了Tax诱导的显著HMGB1启动子活性。这些发现表明,Tax可能通过与C/EBP相互作用,在转录水平上增强HMGB1基因的表达。