亚砷酸钠对人正常肝细胞L-02细胞周期蛋白D1和细胞周期依赖激酶4的影响  

Effects of sodium arsenite on Cyclin D1 and Cyclin dependent kinase 4 of human normal liver cells

作　　者：胡婷[1] 张开菊[2] 张爱华[1] 罗鹏[1] Hu Ting Zhang Kaiju Zhang Aihua Luo Peng(The Key Laboratory of Environmental Pollution Monitoring and Disease Control, Ministry of Education, Department of Toxicology, Guizhou Medical University, Guiyang 550025, China （Hu T, Zhang AH, Luo P Department of Public Health, Guiyang Centre for Disease Control and Prevention, Guiyang 550003, China （Zhang K J)

机构地区：[1]贵州医科大学环境污染与疾病监控教育部重点实验室、卫生毒理学教研室,贵阳550025 [2]贵阳市疾病预防控制中心公卫科,贵阳550003

出　　处：《中华地方病学杂志》2017年第3期191-195,共5页Chinese Journal of Endemiology

基　　金：贵州省科技厅农业攻关计划项目（黔科合NY字[2011]3055号）;贵州省教育厅基金项目（黔省专合字[2010]39号）;贵州省科技厅（黔科合J字[2010]2188）

摘　　要：目的研究亚砷酸钠（NaAsO2）对人正常肝细胞L-02细胞周期蛋白D1（CyclinD1）和细胞周期依赖激酶4（CDK4）表达的影响。方法以0（对照）、50、100、150μmoL／LNaAsO2处理L-02细胞24h，采用流式细胞仪检测细胞周期变化，采用实时荧光定量PCR和蛋白免疫印迹检测Cyclin D1、CDK4mRNA和蛋白表达水平。结果细胞周期检测：与对照组G0．G1期[（60．33±0．40）％]比较，除50μmol／LNaAsO2组细胞比例[（54．58±0140）％]减少外，100、150μmol／LNaAsO2组细胞比例[（64．60±0．62）％、（83．13±0．25）％]显著增加，差异均有统计学意义（P均〈0．05）；对照组，50、100、150μmol／LNaAsO2组S期细胞比例逐渐减少[（34．35±0.30）％、（29．89±0.32）％、（29．50±0.44）％、（11．93±0．12）％]，差异均有统计学意义（P均〈0．05）；对照组，50、100、150μmol／LNaAsO2组G2-M期细胞比例[（5．32±0.11）％、（15．54±0．14）％、（5．90±0．20）％、（4．93±0．15）％]比较，差异有统计学意义（F=908．359，P〈0.05）。CyclinD1和CDK4检测：对照组，50、100、150μmoVLNaAsO2组CyclinD1mRNA（0．48±0．17、1．00±0．31、1．00±0．21、2．06±0．53）和蛋白（0．65±0．02、0．64±0．05、0．71±0．10、0．84±0．05）表达水平比较，差异均有统计学意义（F=167．886、46．575，P均〈0．05）；对照组，50、100、150μmoL／LNaAsO2组CDK4mRNA（1．04±0．19、1.00±0．21、1．29±0．22、2．31±0．31）和蛋白（0．67±0.04、0．74±0．03、0．83±0．07、0．94±0．04）表达水平比较，差异均有统计学意义（F=95．171、145．848，P均〈0．05）。结论NaAsO2通过改变细胞周期调控因子Cyelin D1、CDK4mRNA和蛋白表达水平。使L-02肝细胞的生长周期阻滞于G0—G1期，最终导致细胞损害的发生。Objective To investigate the effects of sodium arsenite （NaAsO2） on the expressions of Cyclin D1 and Cyclin dependent kinase 4 （CDK4） in human normal liver cells （L-02）. Methods L-02 cells were exposed to different doses of NaAsO2 （0, 50, 100, 150 μmol/L） for 24 h, flow cytometry was used to detect the cell cycle, real time quantitative PCR and Western blotting were used to detect the Cyclin D1, CDK4 mRNA and protein expression. Results Cell cycle detection： compared with the control group G0-G1 phase [（60.33 ± 0.40）%], except 50 μmol/L NaAsO2 group [（54.58 ± 0.40）%], the numbers of cells in 100 and 150 μmol/L NaAsO2 groups [（64.60 ± 0.62）%, （83.13 ± 0.25）%] were increased, the differences were statistically significant （all P 〈 0.05）; cell proportion of S phase in the control group, 50, 100 and 150 μmol/L NaAsO2 groups [（34.35 ± 0.30）%, （29.89 ± 0.32）%, （29.50 ± 0.44）%, （11.93 ± 0.12）%] were decreased, the differences were statistically significant （all P 〈 0.05）; cell proportion of G2-M phase was compared between the control group, 50, 100 and 150 p.mol/L NaAs02 groups [（5.32 ± 0.11）%, （15.54 ± 0.14）%, （5.90 ± 0.20）%, （4.93 ± 0.15）%], the difference was statistically significant （F = 908.359, P 〈 0.05）. Cyclin D1 and CDK4 detection： the expressions of Cyclin D1 mRNA （0.48 ± 0.17, 1.00 ± 0.31, 1.00 ± 0.21, 2.06 ± 0.53） and protein （0.65 ± 0.02, 0.64 ± 0.05, 0.71 ± 0.10, 0.84 ± 0.05） were compared betwee the control group, 50, 100 and 150 μmol/L NaAsO2 groups, the differences were statistically significant （F = 167.886, 46.575, all P 〈 0.05）; the expressions ofCDK4 mRNA （1.04 ± 0.19, 1.00 ± 0.21, 1.29 ± 0.22, 2.31 ± 0.31） and protein （0.67 ± 0.04, 0.74 ± 0.03, 0.83 ± 0.07, 0.94 ± 0.04） were compared betwee the control group, 50, 100 and 150 μmol/L NaAsO2 groups, the differences were statistically significant （F = 95.171, 145.848, all P 〈 0.05）. Conclusions Treatment o

关 键 词：亚砷酸盐类 L-02细胞 细胞周期 细胞周期蛋白D1 细胞周期依赖激酶4 

分 类 号：R599.1[医药卫生—内科学]