Polymorphisms of KDR Gene Are Associated With Coronary Heart Disease
Yibo Wang, PhD*,
Yi Zheng, MD*,
Weili Zhang, PhD*,
Hui Yu, MS*,
Kejia Lou*,
Yu Zhang*,
Qin Qin, MD ,
Bingrang Zhao, MD,
Ying Yang, MD and
Rutai Hui, MD, PhD*,*
* Key Laboratory for Clinical Cardiovascular Genetics, Ministry of Education, China and Sino-German Laboratory for Molecular Medicine, and the Department of Cardiology, Cardiovascular Institute and FuWai Hospital, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China
Tianjin Cardiovascular Institute and Tianjin Chest Hospital, Tianjin, China
Qingdao FuWai Hospital, Shandong, China.
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Figure 1 SNP-604C–Bearing KDR Promoter Had Lower Transcription Activity
The pGL3 luciferase reporter contained either the T (pGL3-T) or C allele (pGL3-C) at the promoter –604 locus. Values represent the average of 6 experiments and the bars represent the standard deviation. The pGL3-basic was used as a negative control without any promoter sequence, and pGL3-control as a positive control. KDR = kinase insert domain-containing receptor; SNP = single nucleotide polymorphism.
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Figure 2 SNP1192 and SNP1719 Influenced the Binding Efficiency of VEGF to KDR
HEK293s cells were transfected with 8 µg of pcDNA3.1-KDR (KDR-VH, KDR-IH, KDR-VQ, or KDR-IQ). After 36 h, the cells were rinsed with cold phosphate-buffered saline 3 times, and the binding of vascular endothelial growth factor (VEGF)165 (10 ng/ml, R&D Systems, Minneapolis, Minnesota) was carried out in binding buffer containing DMEM, 25 mmol/l HEPES (pH 7.4), 1 µg/ml heparin, and 0.1% gelatin for 2 h at 4°C. Then the cells were rinsed with cold phosphate-buffered saline 5 times, lysed with cell lysis buffer, followed by enzyme-linked immunosorbent assay. The values were the ratio of VEGF to KDR. The experiments were repeated 3 times, and 2 replicates were performed for each experiment. The values were presented as means, and the T bars represented standard deviations. Abbreviations as in Figure 1.
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Figure 3 Serum Levels of KDR Antigen Were Correlated With Genotypes of SNP-604
Levels of the serum KDR were presented as means, and the T bars represented standard deviations. The correlation was significant (p = 0.013), and Spearman coefficient for the existing correlation was rs = –0.374. Nineteen samples with TT genotype, 14 with TC genotype, and 10 with CC genotype were analyzed. Abbreviations as in Figure 1.
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