内质网应激在氟化物诱导人肝癌细胞HepG2凋亡机制中的作用  

The pathological role of endoplasmic reticulum stress in fluorosis-induced apoptosis of human hepatocellular carcinoma cell line （HepG2）

作　　者：刘咏妍[1] 禹文峰[1] 官志忠[2] 

机构地区：[1] 贵阳医学院分子生物学重点实验室,550004 [2] 550004 贵阳医学院分子生物学重点实验室 550004 贵阳医学院病理学教研室

出　　处：《中华地方病学杂志》2015年第5期331-334,共4页Chinese Journal of Endemiology

基　　金：国家自然科学基金（81160335）;科技部项目（2013BA105803）;贵州省科技厅国际合作项目（黔科合外G字[2011]7014号）

摘　　要：目的 探讨内质网应激在氟化物诱导人肝癌细胞HepG2凋亡机制中的作用.方法 体外培养HepG2细胞24h,分别采用1、3、6、9 mmol/L氟化物（NaF）处理细胞24h,对照组细胞正常培养.流式细胞仪检测各组细胞凋亡情况;提取3 mmol/L NaF处理组细胞RNA和蛋白,实时荧光定量PCR和蛋白质免疫印迹方法检测HepG2细胞中内质网应激反应标志物葡萄糖调节蛋白GRP78、GRP94和CCAAT/增强子结合蛋白同源蛋白（CHOP） mRNA和蛋白的表达.结果 NaF处理HepG2细胞24 h,对照组及1、3、6、9 mmol/L NaF处理组细胞凋亡率分别为（6.25±1.27）％、（13.48±1.00）％、（24.08±1.88）％、（30.19±3.07）％、（37.72±4.43）％,各组间细胞凋亡率比较差异有统计学意义（F=65.828,P＜0.01）.3 mmol/L NaF处理HepG2细胞24 h,GRP78、GRP94和CHOP mRNA表达[（1 172.41±459.60）％、（946.95±635.85）％、（7 846.97±1 670.01）％]均高于对照组[（100.00±1.77）％、（100.00±2.08）％、（100.00±0.74）％,t=12.77、4.67、11.50,P均＜0.01];GRP78和CHOP蛋白表达[（159.99±67.59）％、（155.15±94.24）％]均高于对照组[（100.00±30.68）％、（100.00±41.44）％,t=-3.27、-1.99,P均＜0.05],GRP94蛋白表达[（46.40±41.46）％]低于对照组[（100.00±68.86）％,t=4.02,P＜0.05].结论 内质网应激反应参与了氟化物诱导HepG2细胞凋亡发生的病理过程.Objective To investigate the pathological role of endoplasmic reticulum stress in fluomsisinduced apoptosis in human hepatocellular carcinoma cell line （HepG2）.Methods Under stimulation of 1,3,6,9 mmol/L concentrations of NaF in vitro for 24 h,while normal control group was cultured under normal condition,the apoptosis of HepG2 cells was measured by flow cytometry.The endoplasmic reticulum stress markers （glucose regulative proteins 78,94;GRP78,GRP94） and CCAAT/enhancer-binding protein homologous protein （CHOP） in HepG2 cells were measured at both mRNA and protein levels by real-time PCR and Western blotting,respectively.Results After treated with 0,1,3,6,9 mmol/L NaF for 24 h,the apoptosis rate of HepG2 cells was （6.25 ± 1.27）%,（13.48 ± 1.00）%,（24.08 ± 1.88）%,（30.19 ± 3.07）% and （37.72 ± 4.43）%,respectively,and the difference was statistically significant among groups （F =65.828,P 〈 0.01）.After treated with 3 mmol/L NaF for 24 h,the mRNA level of GRP78,GRP94 and CHOP was （1 172.41 ± 459.60）%,（946.95 ± 635.85）% and （7 846.97 ± 1 670.01）%,which was increased compared to those of the control groups [（100.00 ± 1.77）%,（100.00 ± 2.08）%,（100.00 ± 0.74）%,t =12.77,4.67,11.50,all P 〈 0.01].Under the same condition,the protein levels of GRP78 and CHOP were （159.99 ± 67.59）% and （155.15 ± 94.24）%,which were increased compared to those of the control groups [（100.00 ± 30.68）%,（100.00 ± 41.44）%,t =-3.27,-1.99,all P 〈 0.05],while GRP94 protein level [（46.40 ± 41.46）%] was decreased compared to that of the control group [（100.00 ± 68.86）%,t =4.02,P 〈 0.05].Conclusion Endoplasmic reticulum stress may be involved in NaF-induced cell death in HepG2 cells.

关 键 词：应激 氟化物 细胞凋亡 

分 类 号：R735.7[医药卫生—肿瘤]