营养缺乏对人肺癌细胞自噬的诱导作用  

Starvation-induced autophagy in cultured non-small cell lung cancer cells

作　　者：郭倩倩[1] 刘志燕[2] 姜丽丽[1] 胡婷华[1] 李东繁[2] 刘莹[1] 刘梦洁[1] 梁璇[1] 南克俊[1] 

机构地区：[1]西安交通大学医学院第一附属医院肿瘤内科,陕西西安710061 [2]西安市中心医院呼吸内科,陕西西安710003

出　　处：《南方医科大学学报》2014年第5期627-630,共4页Journal of Southern Medical University

基　　金：国家自然科学基金(81101777);陕西省社发攻关计划(2013k12-08-3)~~

摘　　要：目的观察营养缺乏对人非小细胞肺癌A549及95D细胞自噬活性的影响,建立A549及95D细胞的自噬模型。方法以EBSS缓冲液代替1640完全培养基,饥饿诱导处于对数生长期的A549及95D细胞0、1、2、3、4、5 h后,采用单丹磺酰戊二胺(MDC)荧光染色法检测细胞内自噬泡的形成,并采用Western blot方法检测自噬特异性基因微管相关蛋白1轻链3(LC3)和自噬相关基因Beclin1的蛋白表达水平。结果饥饿处理A549及95D细胞后,胞内MDC荧光颗粒逐渐增多,饥饿4 h达峰值;Beclin1的表达及LC3-II与LC3-I蛋白表达量的比值(LC3-II/LC3-I)随着饥饿时间的延长逐渐增加,分别在饥饿3 h和4 h达峰值。结论营养缺乏可以诱导增强A549及95D细胞的自噬活性,在饥饿4 h时细胞自噬水平达峰值;成功构建了营养缺乏诱导的人肺癌细胞自噬模型,为深入研究自噬在非小细胞肺癌发生发展过程中的作用及其机制奠定了良好的基础。Objective To observe autophagy induced by starvation in non-small cell lung cancer A459 and 95D cells. Methods A549 and 95D cells in logarithmic growth in 1640 medium were cultured in Earle＆#39;s balanced salt solution （EBSS） for 0, 1, 2, 3, 4 or 5 h. Autophagosome formation in the cell culture was observed by MDC fluorescent staining, and the expression of microtubule-associated protein 1 light chain 3 （LC3） and Beclin1 in the cells were detected using Western blotting. Results Compared with the control cells, the cells with prolonged starvation showed increased MDC- positive cells and autophagosome formation. The expression of Beclin-1 and the LC3-II/LC3-I ratio also increased as the starvation prolonged, reaching the peak levels at 3 h and 4 h, respectively. Conclusion Autophagy can be induced by starvation in A549 and 95D cells in correlation with the expression of autophagy-related proteins LC3 and Beclin-1. These cell models of nutritional deficiency-induced autophagy may allow for a better understanding of the role of autophagy in the development of non-small cell lung cancer.

关 键 词：非小细胞肺癌 营养缺乏 自噬 LC3 BECLIN1 

分 类 号：R734.2[医药卫生—肿瘤]