AEG-1对外周T细胞淋巴瘤恶性生物学行为的影响及机制研究

Peripheral T-cell lymphoma is a high- incidence malignant disease, which is considered to be highly aggressive with high death rate. At resent, the pathogenesis of this disease is still unclear. AEG-1 is a newly identidied oncogene. Recent studies have implicated that AEG-1 plays an important role in the biological processes of many cancers. However the molecular mechanism of AEG-1 on peripheral T cell lymphoma remains unclear. Our previous studies show that AEG-1 is overexpressed in peripheral T-cell lymphoma. Patients with high AEG-1 expression had significantly poor overall survival and disease-free survival. In this application, we are attempting to investigate: ①the expression of AEG-1 in normal lymph nodes, peripheral T cell lymphoma tissues and T lymphoma cell lines. ②the malignant biological characteristics and mechanisms of peripheral T-cell lymphoma regulated by abberant AEG-1 expression via in vivo and in vitro studies..③the gene-expression profile related to proliferation, apoptosis, cell cycle, drug resistance and invasion under different AEG-1 expression using digital gene expression technique, which will be identified by Real-time PCR and Western blotting. This study will contribute to a better understanding of the AEG-1 gene and its targets, as well in understanding the role of this gene in T cell lymphoma occurrence and development, and will provide a new insight that targeting AEG-1 may be a promising therapeutic strategy for peripheral T cell lymphoma patients.

外周T细胞淋巴瘤治疗效果差，预后不良，目前发病机制不明。AEG-1是新近发现的一种癌基因，与多种肿瘤发生发展相关。该基因对外周T细胞淋巴瘤的影响如何？其机制是什么？国内外至今未见相关文献报道。本课题组前期研究发现：AEG-1在外周T淋巴瘤中高表达，与化疗敏感性、生存期、预后等临床指标有相关性。本项目拟在前期工作基础上：①进一步检测AEG-1在正常淋巴结组织和外周T细胞淋巴瘤组织及T淋巴瘤细胞系的表达情况；确定AEG-1的表达与外周T细胞淋巴瘤临床指标之间的关系。②通过体内、体外实验阐明AEG-1对外周T细胞淋巴瘤恶性生物学特征的影响及机制。③应用高通量技术检测不同表达水平AEG-1的T淋巴瘤细胞系与增殖、凋亡、耐药、细胞周期等相关的差异表达基因。本研究结果将可能阐明AEG-1在外周T细胞淋巴瘤中的作用和分子机制，并有望发现新的外周T细胞淋巴瘤预后标志和治疗靶点。

本课题组通过进行检测了 AEG-1 基因在 T 细胞淋巴瘤组织中的表达情况及对T 细胞淋巴瘤细胞系(Jurkat 和 Hut-78)的影响，获得了重要的前期实验结果。应用 q RT-PCR、Western-blot 和数字基因表达谱技术等方法检测 AEG-1基因在外周 T 细胞淋巴瘤发生、发展中的作用，经进一步验证靶基因后，最终明确 AEG-1 基因的作用及机制，然后采用蛋白印迹、RT-PCR 和免疫组织化学技术, 对常氧和缺氧条件下 T-NHL细胞 (Hut-78 和 Jurkat 细胞) 中 AEG-1, LC3-II, 和 Beclin-1 的表达进行了蛋白印迹分析。此外, 用 MTT 和 Annexin-V fitcpi 染色法评价了不同浓度的阿霉素 (ADM) 暴露于不同浓度的 ht-78 细胞常氧和缺氧条件下的增殖和凋亡情况。最后, 用MTT 和Annexin-V FITC/PI 染色法评估了AEG-1 对缺氧时暴露于 阿霉素的 Hut-78 细胞的影响, 并进一步利用 3-MA(自体吞噬抑制剂) 确定了其潜在机制。本研究显示，AEG-1通过调节自噬作用与低氧诱导的T-NHL化疗耐药有关，揭示了一种针对缺氧诱导的 T-NHL化学耐药性的新靶点。项目资助共发表了SCI两篇，项目组支出经费14.16万元，剩余经费7.44万元。剩余经费计划用于本项目的后续支出。