亚砷酸钠及代谢物对16HBE细胞c-Myc mRNA及蛋白表达的影响  

Effects of sodium arsenite and its metabolites on the expression of c-Myc mRNA and protein in 16HBE cells

作　　者：马和 刘娜 江金鋆 赵瑞欢 蒋成兰 漆骏[2] 何越峰 MA He;LIU Na;JIANG Jin-jun;ZHAO Rui-huan;JIANG Cheng-lan;QI Jun;HE Yue-feng(Key Laboratory of Public Health and Disease Control,Education Department of Yunnan Province and School of Public Health,Kunming Medical University,Kunming,Yunnan 650500,China;Yunnan Provincial Center for Disease Control and Prevention,Kunming,Yunnan 650500,China)

机构地区：[1]云南省教育厅公共卫生与疾病防控重点实验室暨昆明医科大学公共卫生学院,云南昆明650500 [2]云南省疾病预防控制中心,云南昆明650500

出　　处：《毒理学杂志》2024年第5期367-372,共6页Journal of Toxicology

基　　金：国家自然科学基金(82160607);云南省科技厅昆明医科大学应用基础研究联合专项(202101AY070001-05);昆明医科大学环境与职业砷暴露致癌研究科技创新团队(k1322127)

摘　　要：目的 探究亚砷酸钠及其代谢产物一甲基胂酸(methanearsonic acid,MMA)和二甲基胂酸(dimethylarsenic acid,DMA)在16HBE细胞中对c-Myc mRNA及蛋白表达的影响。方法 将16HBE细胞接种至6孔板,贴壁后,随机分为不同浓度的亚砷酸钠染毒组(0、4、8和12μmol/L)和不同砷化合物染毒组(0、12μmol/L MMA、12μmol/L DMA和12μmol/L NaAsO_(2))染毒48 h。收集细胞进行实时荧光定量PCR法(qRT-PCR)检测c-Myc mRNA表达、蛋白免疫印迹法检测其蛋白及磷酸化蛋白表达、免疫共沉淀(CO-IP)检测其泛素化变化。结果 与对照组比较,不同浓度的亚砷酸钠染毒后c-Myc的mRNA表达下降,其蛋白及磷酸化蛋白表达上升;0、4、8和12μmol/L亚砷酸钠染毒后mRNA表达量分别为1.00±0.015、0.95±0.009、0.56±0.017和0.25±0.003,差异具有统计学意义(LSD-t=2.70、19.19、49.18,P<0.05);对照组、12μmol/L MMA、12μmol/L DMA和12μmol/L NaAsO_(2)染毒后mRNA表达量分别为1.00±0.040、0.65±0.020、0.93±0.028和0.15±0.004,与对照组相比,MMA组具有统计学意义(P<0.05),DMA组则无统计学意义;MMA、DMA均诱导c-Myc的蛋白表达上升,差异均具有统计学意义(LSD-t=7.85、1.42、21.33,P<0.05),而对其磷酸化蛋白表达无明显影响;与对照组相比,c-Myc泛素化水平表达明显下降,泛素化位点K48表达明显下降。结论 亚砷酸钠影响c-Myc的mRNA及蛋白表达,c-Myc的磷酸化表达上升而泛素化降解下降,但其代谢产物MMA、DMA则与亚砷酸钠不同。Objective To investigate the effects of sodium arsenite and its metabolites methanearsonic acid(MMA)and dimethylarsenic acid(DMA)on the expression of c-Myc mRNA and protein in 16HBE cells.Methods 16HBE cells were inoculated into six-well plates and then randomly divided into different concentrations of sodium arsenite exposure groups(O,4,8,12μmol/L)and arsenic compound exposure groups(0,12μmol/L MMA,12μmol/L DMA,12μmol/L NaAsO_(2))for 48 h.The cells were collected for real-time fluorescence quantitative PCR(qRT-PCR)to detect c-Myc mRNA expression,western blotting to detect protein and phosphorylated protein expression,and co-immunoprecipitation(CO-IP)to detect ubiquitination changes.Results The mRNA expression of c-Myc decreased,the protein and phosphorylated protein expression increased after exposure to different concentrations of sodium arsenic,At exposures of O.4,0.8,and 1.2μmol/L NaAs0_(2),the mRNA expression levels were 1.00±0.015,0.95±0.009,and 0.56±0.017,respectively,the differences were statistically significant(LSD-t=2.70,19.19,49.18,P<0.05).In the control group,the mRNA expression level was 1.00+0.040.In the 12μmol/L MMA,12μmol/L DMA,and 12μmol/L NaAs0,groups,the mRNA expression levels were 0.65±0.020,0.93±0.028,and 0.15±0.004,respectively.Compared with the control group,the MMA group presented a statistically significant difference(P<0.05),while the DMA group did not;Both MMA and DMA induced the increase of c-Myc protein expression,with statistical significance(LSD-t=7.85,1.42,21.33,P<0.05),but had no significant effect on phosphorylated protein expression;Compared with the control group,ubiquidized UB expression and K48 ubiquidization decreased significantly.Conclusion Sodium arsenic affects the expression of c-Myc mRNA and protein.,phosphorylation of c-Myc increased and ubiquitination degradation decreased,but its metabolites MMA and DMAweredifferent from sodium arsenite.

关 键 词：亚砷酸钠 C-MYC 泛素化 磷酸化 一甲基胂酸 二甲基胂酸 

分 类 号：R114[医药卫生—卫生毒理学] R99[医药卫生—公共卫生与预防医学]