NPC1L1基因-762T>C多态性与启动子活性的研究

Study on NPC1L1 gene -762T>C polymorphism associated with promoter activity

目的:研究尼曼匹克C1样1(Niemann Pick C1 like 1,NPC1L1)基因启动子-762T>C多态性的活性差异及药物对其的调节作用。方法:分别构建含NPC1L1基因-762T>C多态性的T/C等位基因启动子荧光酶报告基因,测定启动子活性以及不同药物处理后启动子活性的改变。结果:含T或C等位基因的NPC1L1基因启动子活性无统计学差异(P>0.05)。外源性胆固醇和依泽麦布均在高浓度抑制两种启动子活性,辛伐他汀则增强两种启动子活性(P<0.05),但两种基因型启动子之间对药物依泽麦布和辛伐他汀的反应无统计学差异(P>0.05)。结论:胆固醇和依泽麦布能抑制NPC1L1转录活性活性,辛伐他汀则增强NPC1L1转录活性,而-762T>C多态性不影响启动子活性及药物反应。

Objective To investigate the activity of Niemann Pick C1 like 1 （NPC1L1） gene promoter -762T〉C polymorphism the transcriptional activity of -762T〉C polymorphism of Niemann Pick C1 like 1 （NPC1L1） gene and its regulation by drugs. Methods pGL-3 luciferase reporter genes with T and C allele of NPC1L1 gene -762T〉C polymorphism were constructed. The transcriptional activities were measured by dual luciferase reporter assay between the genotypes and after treated with cholesterol, ezetimibe or simvastatin. Results No significant difference of the promoter activity was found between T allele and C allele （P〉0.05）. Both cholesterol and ezetimibe, at higher concertration reduced the promoter activities significantly （P〈0.05）. Conversely, simvastatin increased the promoter activity of NPC1L1 gene （P〈0.05）. However, no statistical difference existed between the two allele in response to either ezetimibe or simvastatin. Conclusions Cholesterol and ezetimibe can inhibit NPC1L1 promoter activity. On contrary, simvastatin enhances NPC1L1 promoter activity. -762T〉C polymorphism has no effect on NPCIL1 promoter activity and response to drugs.