摘要
目的探求长期亚砷酸钠暴露对体外培养的人膀胱上皮细胞（SV-HUC-1）Warburg效应的影响。方法采用体外培养的方法，以0.0（对照）、0.5 μmol/L亚砷酸钠作用SV-HUC-1细胞，分别培养10、20、30周。使用试剂盒分别检测各组SV-HUC-1细胞分泌乳酸和消耗葡萄糖的情况，蛋白质免疫印迹（Western blot）、实时荧光定量PCR（Real-time PCR）检测糖酵解相关基因SCL2A1和己糖激酶（HK2）的蛋白和mRNA表达水平。结果亚砷酸钠处理10、20、30周细胞乳酸分泌水平[（4.67 ± 0.20）、（7.47 ± 0.28）、（12.46 ± 0.47）mmol/L]、葡萄糖消耗水平[（2.86 ± 0.11）、（4.25 ± 0.19）、（6.38 ± 0.05）mmol/L]、HK2蛋白表达水平（1.21 ± 0.06、1.36 ± 0.13、1.60 ± 0.12）均显著高于对照组[（3.04 ± 0.11）、（3.90 ± 0.32）、（4.77 ± 0.24）mmol/L，（2.17 ± 0.15）、（2.48 ± 0.24）、（2.71 ± 0.13）mmol/L，1.00 ± 0.00，P均〈 0.05]。亚砷酸钠处理20、30周细胞SCL2A1、HK2基因mRNA表达水平，SCL2A1蛋白表达水平均显著高于对照组（P均〈 0.05）；处理10周细胞与对照组比较差异无统计学意义（P均〉 0.05）。结论长期低剂量亚砷酸钠暴露可以增强SV-HUC-1细胞糖酵解，诱导Warburg效应发生。
Objective To study the influence of long time exposure to low levels of sodium arsenite on Warburg effect in cultured human epithelial cells （SV-HUC-1） at different times.MethodsSV-HUC-1 cells were exposed to 0.5 μmol/L sodium arsenite for 10, 20, 30 weeks and cells cultured without sodium arsenite for 10, 20, 30 weeks were regarded as control groups in vitro. Lactate assay kit and glucose assay kit were used to measure the lactate secretion and glucose consumption levels, and cells mRNA and protein expressions of SCL2A1 and hexokinase 2 （HK2） were detected using Real-time PCR and Western blotting.ResultsThe levels of lactate secretion [（4.67 ± 0.20）, （7.47 ± 0.28）, （12.46 ± 0.47） mmol/L], glucose consumption [（2.86 ± 0.11）, （4.25 ± 0.19）, （6.38 ± 0.05） mmol/L] and expression of HK2 protein （1.21 ± 0.06, 1.36 ± 0.13, 1.60 ± 0.12） increased significantly after treated with 0.5 μmol/L sodium for 10, 20, 30 weeks compared with those of control groups [（3.04 ± 0.11）, （3.90 ± 0.32）, （4.77 ± 0.24） mmol/L; （2.17 ± 0.15）, （2.48 ± 0.24）, （2.71 ± 0.13） mmol/L; 1.00 ± 0.00; P 〈 0.05]. Compared with control group, the expressions of SCL2A1 mRNA, HK2 mRNA and SCL2A1 protein in SV-HUC-1 cells treated with sodium arsenite for 10 weeks increased but the difference was not statistically significant （P 〉 0.05）. While the expressions of SCL2A1 mRNA, HK2 mRNA and SCL2A1 protein in SV-HUC-1 cells treated with sodium arsenite for 20 and 30 weeks increased significantly compared to those of control groups （P 〈 0.05）.ConclusionLong-term exposure to low concentrations of sodium arsenite can increase glycolysis in SV-HUC-1 and induce Warburg effect.
作者
陈伟
席淑华
Chen Wei;Xi Shuhua(Department of Environmental Health,School of Public Health,China Medical University,Shenyang 110112,China)
出处
《中华地方病学杂志》
CAS
CSCD
北大核心
2018年第10期785-789,共5页
Chinese Journal of Endemiology
基金
国家自然科学基金（81673207）
辽宁省大学生创新创业训练计划资助项目（201710159000097）
