文章摘要
朱 墨,高文涛,钱祝银,苗 毅.DNA甲基化异常对mir-615-5p在胰腺癌细胞株PANC-1中表达影响[J].南京医科大学学报,2011,(1):26~30
DNA甲基化异常对mir-615-5p在胰腺癌细胞株PANC-1中表达影响
Abnormal DNA methylation in pancreatic cancer cell line PANC-1 affects mir-615-5p expression
投稿时间:2010-07-30  
DOI:10.7655
中文关键词: DNA甲基化  miR-615-5p 胰腺癌  5-氮杂胞苷
英文关键词: DNA methylation  miR-615-5p  pancreatic cancer  5-aza-dC
基金项目:江苏省自然科学基金(BK2006241);国家自然科学基金(30500492)
作者单位
朱 墨 南京医科大学第一附属医院胆胰外科,江苏 南京 210029 
高文涛 南京医科大学第一附属医院胆胰外科,江苏 南京 210029 
钱祝银 南京医科大学第一附属医院胆胰外科,江苏 南京 210029 
苗 毅 南京医科大学第一附属医院胆胰外科,江苏 南京 210029 
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中文摘要:
      目的:描述胰腺癌细胞株中DNA异常高甲基化对miRNA表达抑制所起的作用?观察DNA甲基转移酶抑制剂对miR-615-5p在胰腺癌细胞株PANC-1中甲基化状态和表达的影响?方法:甲基化芯片筛选出在胰腺癌细胞株PANC-1中存在异常甲基化的miRNA?采用去甲基化药物5-氮杂胞苷(5-aza-2-adeoxycytidine,5-aza-dC)处理体外培养的PANC-1细胞,甲基化特异性聚合酶链反应(methylation-specific PCR,MSP)结合亚硫酸氢盐修饰结合测序法(bisulfite-sequencing PCR,BSP)检测用药前后miR-615-5p的甲基化状态的改变?采用RT-PCR定量检测用药前后miR-615-5p表达量的差异?结果:通过MSP和BSP发现在胰腺癌细胞株PANC-1中甲基化率明显高于正常组织?对胰腺癌细胞株用去甲基化剂5-aza-dC的作用,发现miR-615-5p甲基化率减少并伴随着表达的重新激活?结论:胰腺癌细胞株PANC-1中的miR-615-5p表达与启动子区甲基化状态有关,启动子区的高甲基化导致PANC-1中miR-615-5p的表达沉默?
英文摘要:
      Objective: To observe the abnormal DNA methylation inhibits the miRNA expression in pancreatic cancer cell line, and DNA methyltransferase inhibitor on the miR-615-5p of the methylation status and expression in the pancreatic cancer cell line PANC-1. Methods: Methylation microarray was used to screen in pancreatic cancer cell line PANC-1 in the abnormal methylation of miRNA. PANC-1 cells were treated with demethylating agent 5-aza-2-adeoxycytidine (5-aza-dC) in vitro, Methylation specific polymerase chain reaction (methyla-tion- specific PCR, MSP) combination of bisulfite modified DNA sequencing method (bisulfite-sequencing PCR, BSP) were used to detect miR-615-5p methylation changes before and after treatment. The changes of miR-615-5p expression was detected by quantitative RT-PCR before and after treatment. Results: Methylation was significantly higher in pancreatic cancer cell line PANC-1 than in normal tissue by MSP and BSP methods. miR-615-5p reduction in methylation was associated with the expression reactivation in the pancreatic cancer cells PANC-1 treated with demethylation agent 5-aza-dC. Conclusion: This study demonstrates that aberrant DNA methylation of the miR-615-5p CpG island is closely linked to their inappropriate silencing in pancreatic cancer cell line PANC-1.
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