[1]杨卫红,闫良,刘利娥,等.双荧光素酶报告基因系统检测CYP3A4*1G增强CYP3A4表达的调控作用[J].郑州大学学报(医学版),2015,(06):765-768.
 YANG Weihong,YAN Liang,LIU Lie,et al.Regulation role of CYP3A4*1G enhancing CYP3A4 expression detected by dual luciferase reporter gene system[J].JOURNAL OF ZHENGZHOU UNIVERSITY(MEDICAL SCIENCES),2015,(06):765-768.
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双荧光素酶报告基因系统检测CYP3A4*1G增强CYP3A4表达的调控作用()
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《郑州大学学报(医学版)》[ISSN:1671-6825/CN:41-1340/R]

卷:
期数:
2015年06期
页码:
765-768
栏目:
论著
出版日期:
2015-12-20

文章信息/Info

Title:
Regulation role of CYP3A4*1G enhancing CYP3A4 expression detected by dual luciferase reporter gene system
作者:
杨卫红1 闫良2 刘利娥3 赵登职4 张卫5 张莉蓉2#
1郑州大学基础医学院法医学系 郑州 4500012郑州大学基础医学院药理学系 郑州 4500013郑州大学公共卫生学院卫生化学与卫生检验系 4500014郑州颐和医院药学部 郑州 4500475郑州大学第一附属医院麻醉科 郑州 450052
Author(s):
YANG Weihong1 YAN Liang2 LIU Li′e3 ZHAO Dengzhi4 ZHANG Wei5 ZHANG Lirong2
1Department of Forensic Medicine,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 4500012Department of Pharmacy,College of Basic Medical Sciences,Zhengzhou University,Zhengzhou 4500013Department of Sanitary Chemistry and Health Inspection,College of Public Health,Zhengzhou University,Zhengzhou 4500014Department of Pharmacy,Yihe Hospital of Zhengzhou,Zhengzhou 4500475Department of Anesthesiology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052
关键词:
CYP3A4*1GCYP3A4启动子双荧光素酶报告基因转录调控
Keywords:
CYP3A4*1GCYP3A4 promoterdual luciferase reporter genetranscriptional regulation
分类号:
R39433/Q756
摘要:
目的:探讨CYP3A4*1G增强CYP3A4基因表达的负调控作用。方法:设计合成包含CYP3A4*1G G或A等位基因的一系列基因片段(外显子10与内含子10交界处287和181 bp),构建CYP3A4*1G正向位于CYP3A4启动子上游的荧光素酶报告基因载体,与内参质粒pRLTK共转染HepG2细胞,通过双荧光素酶报告基因系统检测荧光素酶活性。结果:与CYP3A4启动子相比较, G与A等位基因在287 bp DNA片段中均降低了荧光素酶表达(F=795.575,P<0.001),且G与A等位基因对CYP3A4启动子活性的抑制程度比较差异无统计学意义(P>0.05)。与CYP3A4启动子相比较,G与A等位基因在181 bp DNA片段中均降低了荧光素酶表达(F=23.218,P<0.001),而G与A等位基因对CYP3A4启动子活性的抑制程度比较差异无统计学意义(P>0.05)。结论:在CYP3A4内含子10与外显子10交界处CYP3A4*1G存在与CYP3A4*1G变异无关的CYP3A4启动子的抑制元件,该抑制元件对CYP3A4*1G增强CYP3A4基因表达起负调控作用。
Abstract:
Aim: To study the negative regulation of CYP3A4*1G enhancing CYP3A4 gene expression.Methods: A series of reporter gene vectors carrying CYP3A4*1G G or A allele in different length DNA fragments(exonintron junction,287 or 181 bp) located upstream of the CYP3A4 promoter in the forward direction were constructed. The above recombinant plasmids were cotransfected with internal control plasmid pRLTK into HepG2 cells by LipofectamineTM 2000, and luciferase activity was measured with dual luciferase reporter gene system.Results: Compared with CYP3A4 promoter, the G allele and the A allele in 287 bp DNA fragments both decreased the expression of luciferase(F=795.575,P<0.001),while there was no significant difference in the degree of inhibition to CYP3A4 promoter by the both alleles(P>0.05). Compared with CYP3A4 promoter, the G allele and the A allele in 181 bp DNA fragments both reduced the expression of luciferase(F=23.218, P<0.001), while there was no difference in the extent of inhibition to CYP3A4 promoter by the both alleles(P>0.05).Conclusion: There is a certain inhibition element independent on CYP3A4*1G variation of the CYP3A4 promoter,lying exonintron 10 junction of CYP3A4,and it has negative regulation to CYP3A4*1G enhancing CYP3A4 expression.

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更新日期/Last Update: 1900-01-01