亚砷酸钠诱导的活性氧增加人皮肤细胞N6 -甲基腺苷水平

目的 探究亚砷酸钠诱导的活性氧是否影响N6 -甲基腺苷（m6A）水平，从RNA表观遗传学角度探讨亚砷酸钠的毒性机制。方法 以亚砷酸钠作为活性氧诱导剂，以N -乙酰半胱氨酸作为活性氧清除剂，检测细胞在亚砷酸钠单独处理和亚砷酸钠联合N -乙酰半胱氨酸处理情况下皮肤细胞活性氧积累情况，同时测定m6A及其甲基化酶的mRNA和蛋白水平的改变。结果 在5、10和15 μM的亚砷酸钠处理下，活性氧含量随砷浓度升高而增加；在亚砷酸钠染毒浓度为15 μM时，细胞内总m6A水平水平相比于对照组升高了1.49倍（P＜0.05），m6A甲基化酶（METTL3、METTL14和WTAP）mRNA表达水平相应升高（P＜0.05），METTL14和WTAP蛋白表达水平分别为对照组的1.47倍和1.85倍（P＜0.05）。N -乙酰半胱氨酸可以降低异常升高的活性氧、m6A以及其甲基化酶水平。结论 亚砷酸钠诱导的活性氧能增加人皮肤细胞m6A的修饰水平，其机制可能与上调m6A甲基化酶表达水平有关。

Reactive oxygen species induced by sodium arsenite increases N6-methyladenosine modification in human keratinocyte cells

Objective The aim of this study was to explore whether reactive oxygen species (ROS) induced by sodium arsenite affect N6-methyladenosine(m6A) level, in order to provide a new idea for the study on toxicity induced by arsenite from the perspective of RNA epigenetics. Methods Arsenite was used to induce ROS, and N-acetylcysteine was used to eliminate ROS. ROS level, mRNA and protein levels of m6A and its methylases(METTL3, METTL14, and WTAP) were tested in arsenite treatment group and arsenite combined with N-acetylcysteine treatment group. Results 5, 10 and 15 μM of arsenite elevated ROS level in a concentration-dependent manner. With the treatment of 15 μM arsenite, the m6A level increased 1.49-fold compared to the control (P<0.05), the RNA levels of m6A methylases (METTL3, METTL14 and WTAP) elevated (P<0.05), and the protein levels of METTL14 and WTAP were increased 1.47-fold and 1.85-fold compared to the control, respectively(P<0.05). N-acetylcysteine could suppress abnormal increase of m6A and its methylases levels in HaCaT cells treated by arsenite. Conclusion ROS induced by arsenite increases m6A level possibly via upregulation of m6A methylases.