Department of Oncology, The First Affiliated Hospital, Zhengzhou University, No. 1 Jianshe East Road, Zhengzhou, Henan, 450052, People's Republic of China.
Department of Otorhinolaryngology, The Third Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, 450052, People's Republic of China.
Mol Med. 2018 Jul 9;24(1):35. doi: 10.1186/s10020-018-0033-6.
This study was to examine the link between astrocyte elevated gene-1 (AEG-1) and hypoxia induced-chemoresistance in T-cell non-Hodgkin's lymphoma (T-NHL), as well as the underlying molecular mechanisms.
Expression of AEG-1, LC3-II, and Beclin-1 were initially examined in human T-NHL tissues (n = 30) and normal lymph node tissues (n = 16) using western blot, real-time PCR and immunohistochemistry. Western blot was also performed to analyze the expression of AEG-1, LC3-II, and Beclin-1 in T-NHL cells (Hut-78 and Jurkat cells) under normoxia and hypoxia. Additionally, the proliferation and apoptosis of Hut-78 cells exposed to different concentration of Adriamycin (ADM) in normoxia and hypoxia were evaluated by MTT and Annexin-V FITC/PI staining assay. Finally, the effects of AEG-1 on Hut-78 cells exposed to ADM in hypoxia were assessed by MTT and Annexin-V FITC/PI staining assay, and 3-MA (autophagy inhibitor) was further used to determine the underlying mechanism.
AEG-1, LC3-II and Beclin-1 expression were significantly increased in T-NHL tissues compared with normal tissues. Incubation of Hut-78 and Jurkat cells in hypoxia obviously increased AEG-1, LC3-II and Beclin-1 expression. Hypoxia induced proliferation and reduced apoptosis of Hut-78 cells exposed to ADM. AEG-1 overexpression further increased proliferation and decreased apoptosis of Hut-78 cells exposed to ADM in hypoxia. Moreover, overexpression of AEG-1 significantly inversed 3-MA induced-changes in cell proliferation and apoptosis of Hut-78 cells exposed to ADM in hypoxia.
This study suggested that AEG-1 is associated with hypoxia-induced T-NHL chemoresistance via regulating autophagy, uncovering a novel target against hypoxia-induced T-NHL chemoresistance.
本研究旨在探讨星形细胞上调基因-1（AEG-1）与 T 细胞非霍奇金淋巴瘤（T-NHL）缺氧诱导的化疗耐药之间的关系，并探讨其潜在的分子机制。
采用 Western blot、实时 PCR 和免疫组织化学法检测 30 例 T-NHL 组织和 16 例正常淋巴结组织中 AEG-1、LC3-II 和 Beclin-1 的表达。Western blot 法检测常氧和缺氧条件下 T-NHL 细胞（Hut-78 和 Jurkat 细胞）中 AEG-1、LC3-II 和 Beclin-1 的表达。采用 MTT 和 Annexin-V FITC/PI 染色法检测常氧和缺氧条件下不同浓度阿霉素（ADM）处理 Hut-78 细胞的增殖和凋亡。最后，采用 MTT 和 Annexin-V FITC/PI 染色法评估 AEG-1 对缺氧条件下 ADM 处理的 Hut-78 细胞的影响，并进一步采用 3-MA（自噬抑制剂）确定其潜在机制。
与正常组织相比，T-NHL 组织中 AEG-1、LC3-II 和 Beclin-1 的表达明显增加。常氧和缺氧孵育 Hut-78 和 Jurkat 细胞可明显增加 AEG-1、LC3-II 和 Beclin-1 的表达。缺氧诱导 ADM 处理的 Hut-78 细胞增殖并减少凋亡。AEG-1 过表达进一步增加了缺氧条件下 ADM 处理的 Hut-78 细胞的增殖并减少了凋亡。此外，AEG-1 的过表达显著逆转了 3-MA 诱导的缺氧条件下 ADM 处理的 Hut-78 细胞的增殖和凋亡变化。
本研究表明，AEG-1 通过调节自噬与缺氧诱导的 T-NHL 化疗耐药有关，为针对缺氧诱导的 T-NHL 化疗耐药提供了新的靶点。
