5-Aza-C对人鼻咽癌细胞hTERT基因表达及端粒酶活性的影响

发布时间：2018-03-13 01:24

本文选题：鼻咽癌　切入点：-Aza-C　出处：《肿瘤防治研究》2016年01期 　论文类型：期刊论文

【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)对鼻咽癌细胞亚端粒区D4Z4甲基化状态、h TERT基因表达及端粒酶活性的影响。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;RT-PCR检测h TERT m RNA水平;端粒重复序列扩增法(telomeric repeat amplifi cation protocol,TRAP)法检测端粒酶活性。结果 5-Aza-C处理后,四种鼻咽癌细胞系的亚端粒区D4Z4序列的甲基化水平较处理前明显下降。2.5μmol/L 5-Aza-C处理后,四种鼻咽癌细胞h TERT m RNA的表达明显下调,端粒酶活性显著抑制。结论在鼻咽癌的发生中,亚端粒区的DNA甲基化水平紊乱起了一定的作用,去甲基化药物5-Aza-C能下调h TERT表达,抑制端粒酶活性。探讨鼻咽癌细胞中h TERT表达与亚端粒区Cp G岛甲基化状态之间的关系,可能为亚端粒区染色体结构异常在鼻咽癌发生中的作用提供深刻见解。
[Abstract]:Objective to study the effects of demethylated drug 5-Azacytidineine 5-Aza-Con on the expression of TERT gene and telomerase activity in nasopharyngeal carcinoma (NPC) cell line D4Z4 methylated. Methods Nasopharyngeal carcinoma cells were treated with CNE1CNE2 and 5-Aza-C. The level of h TERT m RNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) of D4Z4 methylation in subtelomere region and telomeric repeat amplifi cation cation assay by telomeric repeat amplifi cation protocol TRAP.Results after 5-Aza-C treatment, the activity of telomerase was detected by telomeric repeat amplifi cation assay. The methylation level of D4Z4 sequence in subtelomere region of four nasopharyngeal carcinoma cell lines decreased significantly after treatment with 2.5 渭 mol/L 5-Aza-C, the expression of h TERT m RNA was down-regulated and telomerase activity was significantly inhibited in the four nasopharyngeal carcinoma cell lines. The demethylation drug 5-Aza-C can down-regulate the expression of h TERT and inhibit telomerase activity. The relationship between the expression of h TERT and the methylation status of subtelomere CpG island in nasopharyngeal carcinoma cells was investigated. It may provide insights into the role of chromosomal abnormalities in subtelomere regions in the development of nasopharyngeal carcinoma.
【作者单位】：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;
【分类号】：R739.63

【相似文献】