摘要
目的:构建肿瘤细胞体外缺糖缺氧模型,观察缺糖缺氧对肿瘤细胞内质网功能和自噬的影响及其机制。方法:连二亚硫酸钠消耗培养基中氧、平衡盐溶液EBSS取代细胞正常培养液构建缺糖缺氧模型;实验分为对照组(Control)、缺糖缺氧4 h组(4 h)、缺糖缺氧8 h组(8 h)、缺糖缺氧12 h组(12 h)。MDC检测细胞内自噬泡的形成;Western Blotting检测各组肿瘤细胞内质网应激-自噬相关蛋白的表达水平。结果:与对照组相比,MDC结果显示细胞内自噬泡数量明显增加;WesternBlotting结果显示,缺糖缺氧各组细胞LC3-Ⅱ表达水平明显升高,同时内质网应激-自噬相关蛋白GRP78、p-eIF2α、p-PERK表达水平也明显升高。结论:缺糖缺氧通过内质网应激-自噬途径诱导Hela细胞发生自噬。
Objective：To investigate the effect of oxygen-glucose deprivation（OGD） in human Hela cell line on the function of endoplasmic reticulum and autophagy. Methods：OGD model was obtained by earle’s balanced salt solution（EBSS） and sodium dithionite.Four groups in the study： control,OGD 4 h,OGD 8 h and OGD 12 h group.To detect the formation of intra-celluar autophgic vacuole by MDC,to detect the expression of proteins related with intra-celluar endoplasmic reticulum-autophagy in each group of human Hela cell line by Western Blotting. Results：Compared with control group,the results of MDC shown the number of intra-celluar autophgic vacuole increased obviously.The results of Western Blotting shown the expression of LC3-Ⅱof each OGD group advanced obviously,while the expression of proteins of GRP78,p-eIF2a,p-PERK related with intra-celluar endoplasmic reticulum-autophagy also increased obviously. Conclusion：Oxygen-glucose deprivation（OGD） induce autophagy of cervical carcinoma Hela cell line by endoplasmic reticulum stress-autophgy pathway.
出处
《中国妇幼保健》
CAS
北大核心
2011年第26期4089-4091,共3页
Maternal and Child Health Care of China
基金
吉林省科技厅资助项目〔201015240〕
关键词
缺糖缺氧
内质网应激
自噬
Oxygen - glucose deprivation （OGD）
Endoplasmic reticulum stress
Autophagy
