Reproductive Medicine Center, The First Affiliated Hospital of SooChow University, SuZhou 215006, China.
Department of Urology, The First Affiliated Hospital of SooChow University, SuZhou 215006, China.
Cell Biol Int. 2018 Sep;42(9):1124-1131. doi: 10.1002/cbin.10971. Epub 2018 Jul 16.
The aim of this study was to investigate the underlying mechanisms of hypoxia-induced apoptosis of GC-2spd (GC-2) cells. The GC-2 cells were treated with or without hypoxia for 12, 24, 48, and 72 h. Apoptosis of GC-2 cells was detected using TUNEL and flow cytometry. Fluorescence microscopic was used to observe the autophagy of GC-2 cells. Hypoxia-inducible factor-1alpha (HIF-1α), apoptosis-related protein and marker protein of autophagy levels were measured by Western blotting. Hypoxia induced apoptosis and autophagy of GC-2 cells, and increased expression of HIF-1α, LC3-II, Beclin-1, and pro-apoptotic protein, but reduced p62 and anti-apoptotic protein level. Meanwhile, hypoxia-induced HIF-1α mediated expression of apoptosis and autophagy-related protein in GC-2 cell. Furthermore, autophagy regulated hypoxia-induced apoptosis of GC-2 cell. Our data suggest that hypoxia induces apoptosis of GC-2 cell by activation of autophagy involving activation of the apoptosis signaling pathway under the hypoxic condition.
本研究旨在探讨缺氧诱导 GC-2spd（GC-2）细胞凋亡的潜在机制。将 GC-2 细胞分别用或不用缺氧处理 12、24、48 和 72 小时。用 TUNEL 和流式细胞术检测 GC-2 细胞的凋亡。荧光显微镜观察 GC-2 细胞的自噬。通过 Western blot 测定缺氧诱导因子-1α（HIF-1α）、凋亡相关蛋白和自噬标志物蛋白的水平。缺氧诱导 GC-2 细胞凋亡和自噬，并增加 HIF-1α、LC3-II、Beclin-1 和促凋亡蛋白的表达，但降低 p62 和抗凋亡蛋白的水平。同时，缺氧诱导的 HIF-1α 介导了 GC-2 细胞中凋亡和自噬相关蛋白的表达。此外，自噬调节了 GC-2 细胞缺氧诱导的凋亡。我们的数据表明，在缺氧条件下，自噬通过激活凋亡信号通路，激活 HIF-1α 诱导 GC-2 细胞凋亡。
