文章摘要
王燕 戴梦源 张自雄 陈始明 肖伯奎 陶泽璋.5-Aza-C 对鼻咽癌细胞端粒长度及细胞增殖的影响[J].,2015,15(36):7004-7008
5-Aza-C 对鼻咽癌细胞端粒长度及细胞增殖的影响
Effect of 5-Aza-C on the Telomere Length and Cell Proliferation ofNasopharyngeal Carcinoma Cell Line
  
DOI:
中文关键词: 鼻咽癌  5-Aza-C  人端粒长度  细胞增殖活性
英文关键词: Nasopharyngeal carcinoma  5-Aza-C  Telomere length  Cell viability
基金项目:国家自然科学基金项目(81001214, 81070766, 81372880);湖北省自然科学基金项目(2012FFB04312)
作者单位
王燕 戴梦源 张自雄 陈始明 肖伯奎 陶泽璋 武汉大学人民医院耳鼻咽喉-头颈外科湖北省恩施市中心医院五官科 
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中文摘要:
      目的:研究去甲基化药物5-氮杂胞嘧啶核苷(5-Azacytidine,5-Aza-C)对鼻咽癌细胞端粒长度及细胞生长增殖的影响。方法: 常规培养鼻咽癌CNE,CNE1,CNE2 及5-8F细胞系,5-Aza-C 处理鼻咽癌细胞后,甲基化测序聚合酶链反应(MSP)法检测亚端粒 区D4Z4 甲基化,端粒限制性片断检测端粒长度,CCK-8 检测细胞增殖。结果:2.5 uM浓度的5-Aza-C处理后,亚端粒区D4Z4 序 列的甲基化水平明显下降,约在15-20%之间,与处理前的甲基化水平(35-48%)有明显差异,差异均有统计学意义(P < 0.05)。在1 uM和2.5 uM浓度的5-Aza-C 处理后,四种细胞的端粒长度明显缩短,长度在2-4.5kb 之间,差异具有统计学意义(P < 0.05)。5uM 的5-Aza-C处理72 h 后,CNE,CNE1,CNE2 和5-8F 的生存率分别为51.27%,50.46%,48.85%,48.83%,10 uM 的5-azaC 处理72 h 后,CNE,CNE1,CNE2 和5-8F的生存率分别为31.64%,32.34%,30.01%,32.10%,与对照组比较差异有统计学意义(P<0.01)。结 论:5-Aza-C 能缩短端粒长度,抑制鼻咽癌细胞生长增殖活性。
英文摘要:
      Objective:To study effect of 5-Azacytidine (5-Aza-C) treatment on telomere length and cell proliferation of nasopharyngeal carcinoma cell line.Methods:CNE, CNE1, CNE2 and 5-8FCell lines were treated for 72 hours with 0, 1, or 2.5 uM of the demethylating agent 5-Aza-C. Subtelomeric level methylation was evaluated with a bisulfate assay, the human telomere length was assayed by Southern blot terminal restriction fragment analysis, the cell viability was detected using a CCK-8 assay.Results:The level of methylation was generally high (35%-48%), but not statistically significant, treatment with 5-Aza-C significantly decreased DNA methylation levels (15%-20%), the degree of demethylation was statistically significant (P < 0.05). The telomere lengths of the four cell lines differed before demethylation treatment, but not remarkably. Compared with the telomere lengths of CNE and CNE1 cells (ranging 2.5-7.5 kb), those of CNE2 and 5-8F cells are highly heterogeneous (ranging 2-9 kb). The telomere lengths of cells that had been treated with 1 uM and 2.5 uM 5-aza-dC were significantly shorter (ranging 2-4.5 kb) compared with untreated cells (P < 0.05). However, there was no statistically significant difference between the two concentrations (P > 0.05). 5-Aza-C inhibited the growth of cell lines in a dose-dependent manner,the survival rate was about 51.27%, 50.46%, 48.85%, 48.83%respectively in the CNE, CNE1, CNE2, and 5-8F cells treated with 5 uM5-azaC (P<0.01) and about 31.64%, 32.34%, 30.01%, 32.10%in the cells after 72 h treated with 10 uM5-AzaC (P<0.01).Conclusion:Our results suggest that 5-Aza-C remarkably shortened telomeres, and effectively inhibit the cell proliferation in NPC cells.
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