缺糖缺氧对经内质网应激途径诱导Hela细胞凋亡的影响  

Effect of oxygen-glucose deprivation on apoptosis of cervical carcinoma cell line Hela induced by endoplasmic reticulum stress pathway

作　　者：徐冶[1,2] 钟加滕[1] 李晓宁[1] 于慧美[1] 康劲松[1] 苏静[1] 孙连坤[1] 

机构地区：[1]吉林大学白求恩医学院病理生理学教研室,吉林长春130021 [2]吉林医药学院医学科研实验室,吉林吉林132013

出　　处：《吉林大学学报（医学版）》2011年第5期821-824,I0002,共5页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅科研基金资助课题(201015240)

摘　　要：目的:体外构建肿瘤细胞缺糖缺氧(OGD)模型,观察缺糖缺氧应激条件对肿瘤细胞凋亡的影响及其机制。方法:利用平衡盐溶液EBSS取代细胞正常培养液构建缺糖模型,利用连二亚硫酸钠消耗培养基中氧构建缺氧模型;实验分为对照组、缺糖缺氧4、8和12 h组。MTT法检测各组细胞增殖率;Western blotting检测各组Hela细胞内质网应激和凋亡相关蛋白的表达水平;Hoechst染色检测各组细胞细胞核形态学改变。结果:与对照组比较,缺糖缺氧各组细胞增殖率明显下降(P<0.05);内质网应激相关蛋白GRP78表达水平明显升高,内质网应激相关凋亡信号蛋白CHOP和caspase-4活化片段蛋白表达水平明显升高(P<0.05);细胞内凋亡相关蛋白caspase-3活化片段表达水平(P<0.05);缺糖缺氧组细胞核出现核皱缩和核碎裂等凋亡形态学表现。结论:缺糖缺氧可以诱导Hela细胞发生凋亡,其凋亡可能是通过内质网应激介导。Objective To construct oxygen-glucose deprivation（OGD） model of human Hela cell line and to investigate the effect of OGD on the apoptosis of tumor cells and its mechanism in vitro.Methods The OGD models were constructed by Earle＇s balanced salt solution（EBSS） and sodium dithionite.Four groups were established such as control group,OGD 4 h group,OGD 8 h group and OGD 12 h group.The changes of proliferation rates of cells in different groups were detected by MTT.The expressions of proteins related to intra-celluar endoplasmic reticulum stress and apoptosis of Hela cells in each group were detected by Western blotting and the morphological changes of nuclear in each group were detected by Hoechst staining.Results Compared with control group,the proliferation rate in OGD group was decreased obviously,the difference had statistical significance（P0.05）;the expression of GRP78 protein related with endoplasmic reticulum stress was significantly in creased,and the expressions of apoptosis signal protein CHOP and the caspase-4 activated fragment protein were also increased（P0.05）,while the expression of protein of caspase-3 activated fragment related to intra-celluar apoptosis was also increased（P0.05）.The morphological changes of nuclear appeared in OGD group,such as nuclear crimple and karyorrhexis.Conclusion OGD can induce the apoptosis of human Hela cell line and the apoptosis may be mediated by endoplasmic reticulum stress.

关 键 词：缺糖缺氧 内质网应激 凋亡 HELA细胞 

分 类 号：R363.2[医药卫生—病理学]