文章摘要
陈 璇,朱 璐,徐凛峰,汪 悦.miR-335启动子活性鉴定及药物干预[J].南京医科大学学报,2015,(1):22~25
miR-335启动子活性鉴定及药物干预
Identification and intervention of promoter regions of miR-335
投稿时间:2014-05-22  
DOI:10.7655/NYDXBNS20150105
中文关键词: microRNA-335  启动子  罗格列酮  胰岛素
英文关键词: miRNA-335  promoter  rosiglitazone  insulin
基金项目:江苏省高校自然科学基金(12KJD360001)
作者单位
陈 璇 南京中医药大学护理学院,江苏 南京 210023 
朱 璐 南京中医药大学护理学院,江苏 南京 210023 
徐凛峰 南京医科大学第二附属医院普外科,江苏 南京 210011 
汪 悦 南京中医药大学第一临床医学院,江苏 南京 210023 
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中文摘要:
      目的:探讨miR-335上游片段是否具有转录活性以及罗格列酮和胰岛素对miR-335启动子区域转录活性的影响。方法:利用PCR技术克隆位于MEST内含子中miR-335上游区域的不同片段,采用双荧光素酶报告基因系统鉴定其转录活性。使用1 μmol/L的罗格列酮及不同浓度的胰岛素进行干预并观察转录活性有无变化。结果:不同的miR-335启动子区域转录活性不同,在miR-335上游约600个碱基区域活性最高。未使用药物干预组细胞中,miR-335启动子区域活性的表达与基础值相比差异无统计学意义(P > 0.05);使用1 μmol/L罗格列酮干预后,miR-335启动子区域活性显著升高(P < 0.01),使用10~12 mol/L的胰岛素干预时,启动子活性升高,但当胰岛素浓度上升后,启动子活性变化并没有统计学差异。结论:miR-335上游约600个碱基为启动子关键区域,同时,miR-335有可能通过自身的转录调控机制参与肥胖与胰岛素抵抗的发生发展。
英文摘要:
      Objective:To explore whether miR-335 has its own transcription unit and effects of rosiglitazone and insulin on transcriptional activity of MiR-335 promoter regions. Methods: We cloned different fragments from upstream sequences of miR-335 in MEST intron by PCR. Then, the luciferase activity of these different plasmids was detected by dual-luciferase reporter system. After the stimulation of 1 μmol/L rosiglitazone and insulin of different concentration, the transcriptional activity change of fragments from upstream sequences of miR-335 was examined. Results: Different miR-335 promoter regions had different transcription activity, and the region of about 600 basic groups in miR-335 upstream exhibited the highest activity. The activity of miR-335 promoter in untreated HEK293T cells showed no statistical significance compared to the baseline value. The levels of miR-335 promoter activity showed significant elevation after the intervention of 1 μmol/L rosiglitazone (P < 0.01). The activity of miR-335 promoter was a little elevated with the stimulation of 10-12 mol/L insulin (P < 0.05). Meanwhile, the levels of miR-335 promoter activity had no statistical significance when the concentration of insulin increased. Conclusion: About 600 basic groups in miR-335 upstream were major promoter regions. Meanwhile, miR-335 may play an important role in the development of obesity and insulin resistance via its own transcription mechanism.
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