摘要
通过使用DNA甲基转移酶(DNA methyltransferases,DNMTs)抑制剂5-氮-2'脱氧胞苷(5-aza-2'deoxycytidine,5-Aza-Cd R)和低氧干预神经细胞系NG108-15,旨为揭示5-Aza-Cd R、低氧、低氧预适应对NG108-15细胞增殖和细胞周期影响的相关机制。利用5-Aza-Cd R(10.0μmol/L)孵育NG108-15细胞后进行低氧与低氧预适应处理;使用倒置显微镜观察细胞形态的变化;通过RTCA分析细胞增殖指数;收集细胞后使用流式细胞仪检测各组细胞周期和细胞凋亡的变化情况;利用Real-time PCR检测甲基转移酶DNMTs和细胞周期相关基因在m RNA水平的表达变化情况。结果表明,5-Aza-Cd R和低氧抑制NG108-15细胞增殖,促进细胞早期和晚期凋亡,低氧预适应促进细胞的增殖。然而,5-Aza-Cd R处理后再低氧与低氧预适应,DNMTs和细胞相关周期基因的m RNA转录增加(P<0.05)。10.0μmol/L的5-Aza-Cd R联合低氧处理抑制细胞的增殖,低氧预适应对细胞的增殖有促进作用。
The aim of this study is to explore the mechanisms of 5-aza-2＇deoxycytidine（5-Aza-Cd R）and/or hypoxia and/or hypoxic preconditioning on the proliferation and cycle of NG108-15 cells while using the inhibitor（5-Aza-Cd R）of DNA methyltransferases（DNMTs）to interfere the nerve cell line NG108-15. The 10.0 μmol/L 5-Aza-Cd R was added to cell culture medium, and then cells were treated in hypoxia and/or hypoxic preconditioning. The cell morphology was observed under inverted microscope. The proliferation of NG108-15 was measured using the RTCA. The cell apoptosis and cell cycle were analyzed through flow cytometry analysis. The m RNA levels of DNMTs and cell cycleassociated gene were analyzed by real-time PCR. 5-Aza-Cd R and hypoxia inhibited NG108-15 cells proliferation, promoted cell early and late apoptosis, and hypoxic preconditioning promoted cell proliferation. However, as cells were treated by hypoxic or hypoxic preconditioning after the cell treated with 5-Aza-Cd R, DNMTs and m RNA transcription of cell cycle-associated genes increased. Conclusively, the jointed treatment of 10.0 μM 5-Aza-Cd R with hypoxic inhibited the proliferation of nerve cell line NG108-15, and hypoxic preconditioning had a promoting effect on the cell proliferation.
出处
《生物技术通报》
CAS
CSCD
北大核心
2016年第1期213-219,共7页
Biotechnology Bulletin
基金
国家自然科学基金项目(81460283)
内蒙古自然科学基金项目(2014MS0810)
自治区研究生科研创新项目[S201410127(Y02)]
