机构：[1]Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Sciences Key Lab, Beijing, China首都医科大学附属北京同仁医院临床科室眼科亦庄院区医学视光科[2]Civil Aviation Medical Center, Civil Aviation Administration of China, Beijing, China
Purpose: To elucidate the role of the protein coding laminin 1 (LAMA1) gene in pathological myopia (PM) at the transcriptional level. To achieve this, the binding affinity of single nucleotide polymorphism (SNP) rs2089760located on the LAMA1 promoter geneto human fetal scleral fibroblast (HFSF) nucleoprotein was investigated and its effect on LAMA1 transcriptional initiation activity was analyzed.Methods: Binding interactions of the HFSF nucleoprotein and biotin-labeled SNP rs2089760 probe were investigated by amplifying the LAMA1 promoter gene and performing overlap extension polymerase chain reaction (PCR) to obtain the G/A mutation of LAMA1 SNP rs2089760. Recombinant adenovirus vectors, Ad5f11p-pLAMA1SNPa-Luc2, Ad5f11p-pLAMA1SNPg-Luc2, and Ad5f11p-CMV-RLuc, were constructed. Fluorescence intensity ratios of firefly luciferase (FLuc) and renilla luciferase (RLuc) vectors were measured 48 h after HFSF infection.Results: Both specific and mutant probes banded precisely with HFSF nucleoprotein. The intensity value of the mutant probe was significantly lower than that of the specific probe (p < 0.05). HFSFs were successfully infected by the recombinant adenoviruses. The FLuc/RLuc fluorescence intensity ratio of Ad5f11p-pLAMA1SNPa-Luc2 (0.0238 +/- 0.0009) was significantly lower than that of Ad5f11p-pLAMA1SNPg-Luc2 (0.0281 +/- 0.0015) (p < 0.05).Conclusions: It is highly likely that SNP rs2089760 in the LAMA1 promoter region is located at the transcription factor binding site. The SNP rs2089760 G > A mutation reduces transcription factor binding ability and transcriptional initiation activity, and negatively regulates gene transcription of LAMA1. We suggest that LAMA1 SNP rs2089760 plays an important role in the development of PM.
基金：
Beijing Natural Science FoundationBeijing Natural Science Foundation [7122041]; 215 High level Talent Fund of Beijing Health Government [2013-2-023]; National Natural Science Foundation of ChinaNational Natural Science Foundation of China (NSFC) [81271041]
第一作者机构：[1]Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Sciences Key Lab, Beijing, China[2]Civil Aviation Medical Center, Civil Aviation Administration of China, Beijing, China
通讯作者：
通讯机构：[1]Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Sciences Key Lab, Beijing, China[*1]Beijing Tongren Eye Center, Beijing Tongren Hospital, Capital Medical University, Beijing Ophthalmology & Visual Sciences Key Lab, 1 Dong Jiao Min Xiang, Beijng 100730, China
推荐引用方式(GB/T 7714)：
Liang Yanchuang,Song Yanzheng,Zhang Fengju,et al.Effect of a Single Nucleotide Polymorphism in the LAMA1 Promoter Region on Transcriptional Activity: Implication for Pathological Myopia[J].CURRENT EYE RESEARCH.2016,41(10):1379-1386.doi:10.3109/02713683.2015.1118129.
APA：
Liang, Yanchuang,Song, Yanzheng,Zhang, Fengju,Sun, Mingshen&Wang, Ningli.(2016).Effect of a Single Nucleotide Polymorphism in the LAMA1 Promoter Region on Transcriptional Activity: Implication for Pathological Myopia.CURRENT EYE RESEARCH,41,(10)
MLA：
Liang, Yanchuang,et al."Effect of a Single Nucleotide Polymorphism in the LAMA1 Promoter Region on Transcriptional Activity: Implication for Pathological Myopia".CURRENT EYE RESEARCH 41..10(2016):1379-1386
