维生素D受体基因FokI多态对CYP3A4转录调控的影响

中国临床药理学与治疗学 ›› 2009, Vol. 14 ›› Issue (6): 664-669.

维生素D受体基因FokI多态对CYP3A4转录调控的影响

王果¹, 谢海棠^1,2, 李宝群³, 范岚¹, 钱荣华¹, 胡东莉¹, 李智¹

¹中南大学临床药理研究所, 长沙 410078, 湖南;
²皖南医学院弋矶山医院安徽省药物临床评价中心, 芜湖 241001, 安徽;
³承德医学院药理教研室, 承德 067000, 河北

收稿日期:2009-04-25 修回日期:2009-05-26 出版日期:2009-06-26 发布日期:2020-10-27
通讯作者: 李智, 男, 博士, 讲师, 研究方向:遗传药理与临床药理学。Tel:0731-4805380 E-mail:lizhi489@163.com
作者简介:王果,男,在站博士后,讲师,研究方向:遗传药理与临床药理学。Tel:0731-4805380 E-mail:wangguo30@yahoo.com.cn
基金资助:
中国博士后基金一等资助项目(20070410314);中南大学博士后基金资助项目

Effects of vitamin D receptor Fok I polymorphism on the transcriptional control of CYP3A4

WANG Guo¹, XIE Hai-tang^1,2, LI Bao-qun³, FAN Lan¹, QIAN Rong-hua¹, HU Dong-li¹, LI Zhi¹

¹Institute of Clinical Pharmacology, Central South University, Changsha 410078, Hunan, China;
²YijishanHospital of Wannan Medical College, Anhui Provincial Center for Drug Clinical Evaluation, Wuhu 241001, Anhui, China;
³Pharmacal Department of Chengde Medical College, Chengde 067000, Hebei, China

Received:2009-04-25 Revised:2009-05-26 Online:2009-06-26 Published:2020-10-27

摘要/Abstract

摘要： 目的: 研究人维生素D 受体(human vitamin D receptor, hVDR) 基因翻译起始密码子Fok I 突变介导CYP3A4 诱导表达的调控能力与野生型比较有无统计学差异。方法: 从肝脏组织标本中提取总RNA, RT-PCR 法扩增获得野生型hVDR 基因cDNA 序列, 并用定点诱变的方法获得含Fok I 突变的hVDR cDNA 序列, 构建野生型和突变型的真核表达载体;同时构建含CYP3A4 近端ER6 元件的荧光素酶报告基因载体。将hVDR 真核表达载体、CYP3A4 报告基因载体和内对照载体共同瞬时转染COS-7 细胞, 转染24 h 后用1、10、100 nmol/L的1, 25(OH)₂D₃ 孵育48 h, 最后裂解细胞分析荧光素酶活性。结果: Fok I 突变型和野生型hVDR真核表达载体以及CYP3A4 荧光素酶报告载体均构建成功。瞬转后胞内荧光素酶活性检测显示,各浓度1, 25(OH)₂D₃ 孵育后hVDR 基因Fok I 突变型组与野生型组均可激活CYP3A4 的转录;但各维生素D₃ (vitamin D₃, VD₃) 浓度下, 突变型组与野生型组的荧光素酶比活性值没有统计学差异。结论: 与VD₃ 结合后, hVDR Fok I 突变型与野生型均能激活CYP3A4 的诱导表达, 但与野生型比较,Fok I 突变不能导致VDR 蛋白转录激活调控能力的改变。

关键词: 维生素D 受体, CYP3A4, Fok I 突变, 转录调控

Abstract: AIM: To study whether there is significant difference in the inductive effects on CYP3A4 between human vitamin D Receptor (hVDR) wild type and start codon Fok I mutant. METHODS: Total RNA was extracted from liver samples.cDNA of hVDR wild type was acquired by RT-PCR, and cDNA with hVDR Fok I mutant was acquired using site-directed mutagenesis.Eukaryotic expression vectors of both wild type and mutant of hVDR were constructed according to cDNA.Meanwhile, luciferase reporter gene vector was constructed with proximal ER6 element of CYP3A4. COS-7 cells with hVDR eukaryotic expression vectors, CYP3A4 luciferase reporter gene vector and the internal control vector were transiently transfected for 24 h and then were incubated with 1 nmol/L, 10 nmol/L and 100 nmol/L 1, 25(OH)₂D₃ for 48 h.Finally the cells were lysed and the luciferase activity was detected with cell lysis. RESULTS: hVDR eukaryotic expression vectors of wild type and Fok I mutant, and CYP3A4 luciferase reporter gene vector were successfully constructed.The results of luciferase activity showed that CYP3A4 transcription could be activated by hVDR eukaryotic expression vectors of wild type and Fok I mutant when incubated with 1 nmol/L, 10 nmol/L and 100 nmol/L 1, 25 (OH)₂D₃, respectively.However, there was no significant difference between the luciferease activities of hVDR wild type and mutant. CONCLUSION: Both hVDR wild type and Fok I mutant have inductive effect on CYP3A4 when incubated with vitamin D₃ (VD₃).But Fok I mutant could not significantly change the regulational capacity of wild type VDR on CYP3A4.

Key words: vitamin D receptor, CYP3A4A, Fok I mutation, transcriptional control

中图分类号:

R969

王果, 谢海棠, 李宝群, 范岚, 钱荣华, 胡东莉, 李智. 维生素D受体基因FokI多态对CYP3A4转录调控的影响[J]. 中国临床药理学与治疗学, 2009, 14(6): 664-669.

WANG Guo, XIE Hai-tang, LI Bao-qun, FAN Lan, QIAN Rong-hua, HU Dong-li, LI Zhi. Effects of vitamin D receptor Fok I polymorphism on the transcriptional control of CYP3A4[J]. Chinese Journal of Clinical Pharmacology and Therapeutics, 2009, 14(6): 664-669.

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