5-Aza-CdR对低氧NG108-15细胞的影响机制研究

doi:10.13560/j.cnki.biotech.bull.1985.2016.01.034

生物技术通报 ›› 2016, Vol. 32 ›› Issue (1): 213-219.doi: 10.13560/j.cnki.biotech.bull.1985.2016.01.034

5-Aza-CdR对低氧NG108-15细胞的影响机制研究

张柱霞, 孙明英, 杨洁, 姜树原, 闫少春, 邵国

包头医学院中心实验室生物医学研究中心, 包头 014010

收稿日期:2015-03-16 出版日期:2016-01-09 发布日期:2016-01-22
作者简介:张柱霞, 女, 硕士研究生, 研究方向：生物化学和分子生物学, E-mail：836796250@qq.com;孙明英同为本文第一作者
基金资助:
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)］

The Affecting Mechanism of 5-aza-2'-deoxycytidine on the NG108-15 Cells in Hypoxia

ZHANG Zhu-xia, SUN Ming-ying, YANG Jie, JIANG Shu-yan, YAN Shao-chun, SHAO Guo

Biomedical Research Center of Center Laboratory, Baotou Medical College, Baotou 014010

Received:2015-03-16 Published:2016-01-09 Online:2016-01-22

摘要/Abstract

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-CdR)和低氧干预神经细胞系NG108-15, 旨为揭示5-Aza-CdR、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-CdR(10.0 μmol/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mRNA水平的表达变化情况。结果表明, 5-Aza-CdR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-CdR处理后再低氧与低氧预适应, DNMTs和细胞相关周期基因的mRNA转录增加(P<0.05)。10.0 μmol/L的5-Aza-CdR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。

关键词: NG108-15细胞, 5-氮-2&apos, 脱氧胞苷, 低氧, 低氧预适应

Abstract: The aim of this study is to explore the mechanisms of 5-aza-2'deoxycytidine(5-Aza-CdR)and/or hypoxia and/or hypoxic preconditioning on the proliferation and cycle of NG108-15 cells while using the inhibitor(5-Aza-CdR)of DNA methyltransferases(DNMTs)to interfere the nerve cell line NG108-15. The 10.0 μmol/L 5-Aza-CdR 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 mRNA levels of DNMTs and cell cycle-associated gene were analyzed by real-time PCR. 5-Aza-CdR 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-CdR, DNMTs and mRNA transcription of cell cycle-associated genes increased. Conclusively, the jointed treatment of 10.0 μM 5-Aza-CdR with hypoxic inhibited the proliferation of nerve cell line NG108-15, and hypoxic preconditioning had a promoting effect on the cell proliferation.

Key words: NG108-15 cell line, 5-aza-2'-deoxycytidine, hypoxic, hypoxic preconditioning

张柱霞, 孙明英, 杨洁, 姜树原, 闫少春, 邵国. 5-Aza-CdR对低氧NG108-15细胞的影响机制研究[J]. 生物技术通报, 2016, 32(1): 213-219.

ZHANG Zhu-xia, SUN Ming-ying, YANG Jie, JIANG Shu-yan, YAN Shao-chun, SHAO Guo. The Affecting Mechanism of 5-aza-2'-deoxycytidine on the NG108-15 Cells in Hypoxia[J]. Biotechnology Bulletin, 2016, 32(1): 213-219.

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5-Aza-CdR对低氧NG108-15细胞的影响机制研究

The Affecting Mechanism of 5-aza-2'-deoxycytidine on the NG108-15 Cells in Hypoxia

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