亚砷酸钠及砷代谢产物对A549细胞中核孔蛋白107线状和环状RNA表达的影响

Effects of sodium arsenite and arsenic metabolites on the expression of linear and circular RNA of Nup107 in A549 cells

目的探讨亚砷酸钠及砷甲基化代谢产物一甲基胂酸(MMA)、二甲基胂酸(DMA)对人肺腺癌细胞系A549细胞中核孔蛋白107(Nup107)线状RNA和环状RNA(circRNA)表达的影响。方法(1)取对数生长期A549细胞随机分为4组,分别以浓度为0、30、60、90μmol/L的亚砷酸钠刺激48 h。(2)取对数生长期A549细胞随机分为4组;其中,MMA组、DMA组、亚砷酸钠组分别予浓度为90μmol/L的MMA、DMA和亚砷酸钠刺激48 h;对照组细胞不予染毒处理。培养结束后,收获细胞,采用实时荧光定量聚合酶链式反应检测Nup107线状RNA和circRNA的基因表达。结果(1)随着亚砷酸钠剂量的增加,A549细胞中Lin-Nup107相对表达水平下降(P<0.01),4种cirCRNA异构体hsa_circ_0003599、hsa_circ_0027477、hsa_circ_0027478、hsa_circ_0027479相对表达水平均增加(P值均<0.01),均呈剂量-效应关系。与对照组比较当以浓度为90μmol/L的亚砷酸钠刺激时,A549细胞中Lin-Nup107相对表达水平下降(P<0.01),4种cirCRNA异构体相对表达水平均升高(P值均<0.01)。(2)与对照组比较,A549细胞中Lin-Nup107相对表达水平在MMA组和DMA组均升高(P值均<0.05),在亚砷酸钠组下降(P<0.05)。亚砷酸钠组A549细胞中Lin-Nup107相对表达水平分别低于MMA组和DMA组(P值均<0.05)。亚砷酸钠组A549细胞中hsa_circ_0003599、hsa_circ_0027478、hsa_circ_0027479相对表达水平均高于对照组、MMA组和DMA组(P值均<0.05);DMA组A549细胞中hsa_circ_0027479相对表达水平低于对照组(P<0.05)。结论亚砷酸钠可抑制Nup107线状RNA和促进其circRNA基因在A549细胞中的表达,且存在剂量-效应关系。砷的甲基化代谢产物MMA和DMA可导致A549细胞中Nup107线状RNA表达上调,但对circRNA表达的影响不明显。

Objective To explore the effect of sodium arsenite and arsenic metabolites monomethylarsenic acid(MMA) and dimethylarsenic acid(DMA)on the expression of linear and circularRNAs of nucleoporin 107(Nup107) in human lung adenocarcinoma A549 cells. Methods i) The A549 cells in logarithmic phase were treated with 0, 30, 60, 90 μmol/L sodium arsenite for 48 hours. ii)The A549 cells in logarithmic phase were treated with 90 μmol/L sodium arsenite, MMA and DMA for 48 hours, the control group received no treatment. After culturing, the relative expression of linear RNA and circular RNA(circRNA) of Nup107 was detected by real-time quantitative polymerase chain reaction. Results i) The relative expression of linear RNA of Nup107 decreased and four circRNA isomers such as hsa_circ_0003599, hsa_circ_0027477, hsa_circ_0027478 and hsa_circ_0027479 increased with the increase of sodium arsenite dose(all P<0.01), showing a dose-effect relationship. In the 90 μmol/L sodium arsenite stimulated group, the relative expression of linear RNA of Lin-Nup107 decreased, and the four circRNA isomers increased compared with the control group in the A549 cells(all P<0.01). ii) The relative expression of Lin-Nup107 increased in the MMA and DMA stimulated groups and decreased in the sodium arsenite stimulated group compared with the control group in the A549 cells(all P<0.05). The relative expression of Lin-Nup107 decreased in the sodium arsenite stimulated group compared with the MMA and DMA stimulated groups in the A549 cells(all P<0.05). The relative expressions of hsa_circ_0003599, hsa_circ_0027478, hsa_circ_0027479 in the sodium arsenite stimulated group were higher than that in the MMA and DMA stimulated groups as well as the control group(all P<0.05).The relative expressions of hsa_circ_0027479 in the DMA stimulated group was lower than that in the control group(P<0.05). Conclusion Sodium arsenite can induce the down-regulation of linear RNA and the up-regulation of circRNA of Nup107 in a dose-dependent manner. The metabolites of arsenic MMA and DMA can induce the overexpression of linear RNA of Nup107, however, they had no obvious effect on the circRNA of Nup107 in A549 cells.