摘要
目的:探究5-氮杂-2′-脱氧胞苷(5-Aza-CdR)对小鼠海马神经元细胞系HT22细胞增殖及其对DNA甲基化转移酶(DNMT)表达的影响。方法:5-Aza-CdR处理HT22细胞,采用Real-time PCR和免疫印迹检测DNMT1、DNMT3A、DNMT3BmRNA及蛋白的表达;甲基转移酶活性试剂盒检测DNMTs活性;流式细胞仪对其细胞周期及凋亡进行检测。结果:5-Aza-CdR处理HT22细胞24h后,不同浓度5-Aza-CdR均显著抑制HT22细胞增殖,且使胞质空泡化。5-Aza-CdR降低了HT22细胞早期凋亡,但促进晚期凋亡,并诱导细胞周期发生S期阻滞。DNMT1和DNMT3A的mRNA和蛋白表达下降,但DNMT3B表达未发生明显变化。DNMT1和DNMT3B活性降低。结论:5-Aza-CdR可降低DNMT1和DNMT3A mRNA和蛋白表达以及DNMT1和DNMT3B活性,影响HT22细胞周期及凋亡。
Objective:To investigate the effect of 5-Aza-CdR on the proliferation and the expression of DNMTs of mouse hippocampal neuron cell line HT22.Methods:HT22 cells were treated with 5-Aza-CdR.Real-time PCR and Western blotting were used to detect the expression of DNMT1,DNMT3A and DNMT3B.Methyltransferase activity kit was used to detect DNMTs activity,and flow cytometry was used to detect the cycle and apoptosis of cells.Results:HT22 cells were treated with 5-Aza-CdR for 24h.The results showed that different concentrations of 5-Aza-CdR significantly inhibited the proliferation of HT22 cells and caused cytoplasmic vacuolation.The 5-Aza-CdR treatment reduced the early apoptosis of HT22 cells,but enhanced the late apoptosis and induced S phase arrest in the cell cycle.DNMT1 and DNMT3A mRNA and protein expression decreased,but DNMT3B expression did not change significantly.DNMT1 and DNMT3B activity decreased.Conclusion:5-Aza-CdR can reduce the mRNA and protein expression of DNMT1 and DNMT3A,as well as the activity of DNMT1 and DNMT3B,and affect the cell cycle and apoptosis of HT22 cells.
作者
田小莉
杨洁
杨静
闫少春
时静华
邵国
Tian Xiaoli;Yang Jie;Yang Jing;Yan Shaochun;Shi Jinghua;Shao Guo(Biomedical Research Center of Central Laboratory,Institute of Neuroscience,Baotou Medical College,Baotou014010;Key Laboratory of Hypoxia Translational Medicine of Inner Mongolia Autonomous Region,Baotou014010;Key Laboratory of Hypoxia Adaptation and Translational Medicine,Xuanwu Hospital,Capital Medical University,Beijing100053,China)
出处
《解剖学杂志》
CAS
CSCD
2018年第4期404-408,共5页
Chinese Journal of Anatomy
基金
国家自然科学基金(81160244,81360316,81460283,81660307)
内蒙古自然科学基金(2015MS0322,2016MS(LH)0307)
内蒙古自治区高等学校青年科技英才支持计划资助(NJYT-13-A10)
中国博士后科学基金(20080430851)
教育部留学回国人员科研启动基金(46批)
内蒙古卫生厅项目(201302101)
内蒙古教育厅基金(NJZY13243号)
包头医学院基金项目(BYJJ-DF201602,BYJJ-YF201615,BSJJ201617,BYJJ-QM201633,BYJJ-QM201656,BYJJ201502)
