Neoplasma. 2018;65(1):42-48. doi: 10.4149/neo_2018_161102N528.
To investigate the role of Beclin1 in cisplatin-induced apoptosis in laryngeal carcinoma cells Hep-2 and to explore the potential mechanism. We up-regulated Beclin1 expression in Hep-2 cells. The survival rate and apoptotic rate were evaluated by MTT and flow cytometry (FCM). The Beclin1 overexpression group and the control group were treated with cisplatin for 24 hours. The proliferation and cell apoptosis of laryngeal cancer cell lines were evaluated. The mitochondrial membrane potentials were detected by DiOC6(3). Activities of Caspase-8/9/3 and convention of microtubule-associated protein one light chain 3 (LC3) were detected by western blot. The effect of Bcl-2 overexpression on increased cisplatin-sensitivity and autophagy induced by Beclin1 was investigated using Bcl-2 cDNA transfection. Expression of Beclin1 in Hep-2 cells was meaningfully enhanced by transfection, and the proliferation and the apoptosis were not considerably affected. By cisplatin treatment, the Beclin1 overexpression group showed lower survival rate and higher apoptotic rate than the control group (p<0.05). Decrease of mitochondrial membrane potential and increase of activities of Caspase-8, Caspase-9 and Caspase-3 were detected. Beclin1 overexpression increase the convention of LC3, especially after the cisplatin treatment. Overexpression of Bcl-2 decreased the cisplatin-induced apoptosis and inhibited Beclin1-induced autophagy. In conclusion, Beclin1 enhances cisplatin-induced apoptosis in laryngeal carcinoma cells Hep-2 via Bcl-2 modulated autophagy.
为了研究 Beclin1 在顺铂诱导喉癌细胞 Hep-2 凋亡中的作用，并探讨其潜在机制。我们上调了 Hep-2 细胞中的 Beclin1 表达。通过 MTT 和流式细胞术（FCM）评估细胞存活率和凋亡率。用顺铂处理 Beclin1 过表达组和对照组 24 小时，评估喉癌细胞系的增殖和细胞凋亡。用 DiOC6（3）检测线粒体膜电位。用 Western blot 检测 Caspase-8/9/3 的活性和微管相关蛋白轻链 3（LC3）的转化。用 Bcl-2 cDNA 转染研究 Bcl-2 过表达对 Beclin1 诱导的顺铂敏感性增加和自噬的影响。转染后 Hep-2 细胞中 Beclin1 的表达显著增强，但增殖和凋亡没有明显影响。用顺铂处理后，Beclin1 过表达组的存活率低于对照组，凋亡率高于对照组（p<0.05）。检测到线粒体膜电位降低和 Caspase-8、Caspase-9 和 Caspase-3 活性增加。Beclin1 过表达增加了 LC3 的转化，特别是在用顺铂处理后。Bcl-2 的过表达降低了顺铂诱导的凋亡，并抑制了 Beclin1 诱导的自噬。总之，Beclin1 通过 Bcl-2 调节的自噬增强了喉癌细胞 Hep-2 中顺铂诱导的凋亡。
