›› 2012, Vol. 24 ›› Issue (3): 179-182.doi: 10.3969/j.issn.1004-616x.2012.03.004

• 论著 • 上一篇    下一篇

5氮杂脱氧胞苷对鼻咽癌细胞RASSF1A基因甲基化和mRNA表达的影响

叶 静,李明华,张芳婷,万汇涓,龙 霞   

  1. 北京大学深圳医院中心实验室，广东 深圳 518036
  • 收稿日期:2011-11-30 修回日期:2012-02-05 出版日期:2012-05-30 发布日期:2012-05-30
  • 通讯作者: 叶 静

Effect of 5-Aza-CdR on methylation and mRNA expression of RASSF1A genein nasopharyngeal carcinoma cells

YE Jing，LI Ming-hua，ZHANG Fang-ting，WAN Hui-juan，LONG Xia   

  1. Central Laboratory, Peking University Shenzhen Hospital, Shenzhen 518036, Guangdong, China
  • Received:2011-11-30 Revised:2012-02-05 Online:2012-05-30 Published:2012-05-30
  • Contact: YE Jing

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摘要： 目的： 研究5氮杂脱氧胞苷对鼻咽癌CNE2细胞中RASSF1A基因甲基化和mRNA表达的影响。方法：用不同浓度 (0、5、10、20 μmol/L) 5氮杂脱氧胞苷处理CNE2细胞，采用甲基化特异性PCR (MSP)法对处理后细胞中RASSF1A基因甲基化状态进行检测；并用SYBR Green qReal Time-PCR法检测RASSF1A mRNA的表达。结果：阴性对照组CNE2细胞中，RASSF1A基因呈完全甲基化状态，当用5 μmol/L 5氮杂脱氧胞苷处理后，CNE2细胞出现非甲基化产物，20 μmol/L 5氮杂脱氧胞苷处理后，CNE2细胞甲基化状态完全被逆转，均为非甲基化产物。阴性对照组CNE2细胞RASSF1A基因呈低表达，经5~20 μmol/L 5氮杂脱氧胞苷处理后，RASSF1A mRNA的相对表达量逐渐增加，10和20 μmol/L 5氮杂脱氧胞苷处理组，RASSF1A mRNA表达水平均明显高于阴性对照组 (P<0.01)；且20 μmol/L处理组明显高于5 μmol/L处理组 (P<0.01)。结论：CNE2细胞RASSF1A基因甲基化可以被5氮杂脱氧胞苷逆转，且5氮杂脱氧胞苷可促进RASSF1A mRNA的表达。

关键词: 鼻咽癌, CNE2, RASSF1A, 甲基化, mRNA表达

Abstract: OBJECTIVE: The relationship between methylation of RASSF1A gene and mRNA expression in nasopharyngeal carcinoma cell line CNE2 was investigated. METHODS：CNE2 cells were treated with various concentrations of 5-Aza-CdR. Methylation of RASSF1A gene was evaluated by methylation-specific polymerase chain reaction(MSP). Expression of RASSF1A mRNA was detected by SYBR Green qRealTime-PCR. RESULTS：RASSF1A gene was completely methylated in the negative control group cell. De-methylation occurred in the 5 μmol/L of 5-Aza-CdR. Methylation was completely reversed by 20 μmol/L of 5-Aza-CdR. The expression of RASSF1A mRNA in the negative control group was low. The expression level increased gradually by 5，10 and 20 μmol/L 5-Aza-CdR treatment. The levels in 10 and 20 μmol/L treated group were significantly improved in comparison with the negative control group (P<0.01). RASSF1A mRNA in 20 μmol/L treated group was obviously higher than that in 5 μmol/L treated group (P<0.01). CONCLUSION：Methylation of RASSF1A gene in the nasopharyngeal carcinoma cell line CNE2 could be reversed by 5-Aza-CdR， promoting the expression of RASSF1A gene mRNA.

Key words: nasopharyngeal carcinoma, CNE2, RASSF1A, methylation, expression

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