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Association of COL4A1 genetic polymorphisms with coronary artery disease in Uygur population in Xinjiang, China.

Adi D, Xie X, Ma YT, Fu ZY, Yang YN, Li XM, Xiang Y, Liu F, Chen BD - Lipids Health Dis (2013)

Bottom Line: For total and men, the rs605143 was found to be associated with CAD by in a dominate model (p = 0.014, p = 0.013, respectively).The rs565470 was also found to be associated with CAD in a recessive model for total and men (both p < 0.001), and the difference remained statistically significant after multivariate adjustment (P = 0.002, P = 0.001, respectively).Both rs605143 and rs565470 of COL4A1gene are associated with CAD in Uygur population of China.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, People's Republic of China. myt_xj@sina.com.

ABSTRACT

Background: Type IV collagen is important for the structural integrity and function of basement membranes. Basement membranes surround vascular smooth muscle cells in the media, COL4A1 is the most abundant component of type IV collagen in all Basement membranes. However, the relationship between COL4A1 genetic polymorphisms and coronary artery disease (CAD) remains unclear. We performed a case-control study to explore the association of COL4A1 genetic polymorphisms with CAD in Uygur population of China.

Methods: 1095 Uygur people (727 men, 368 women) including 471 CAD patients and 624 controls were selected for the present study. Two SNPs (rs605143 and rs565470) were genotyped by using the polymerase chain reaction-restriction fragment length (PCR-RFLP) method.

Results: For total and men, the rs605143 was found to be associated with CAD by in a dominate model (p = 0.014, p = 0.013, respectively). The difference remained statistically significant after multivariate adjustment (p = 0.036, p = 0.014, respectively). The rs565470 was also found to be associated with CAD in a recessive model for total and men (both p < 0.001), and the difference remained statistically significant after multivariate adjustment (P = 0.002, P = 0.001, respectively).

Conclusion: Both rs605143 and rs565470 of COL4A1gene are associated with CAD in Uygur population of China.

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Related in: MedlinePlus

Restriction fragment length polymorphism analysis for determination of genotype. A. For SNP1, the GG genotype shows two bands of 362 bp and 220 bp (1 and 2); The AG genotype shows three bands of 362 bp, 258 bp and 220 bp (3, 4, 6 and 7), The AA genotype shows three bands of 258 bp, 220 bp and 104bp (5). B. For SNP2, the TT genotype shows one band of 416 bp (1 and 4); The CT genotype shows two bands of 416 bp and 324 bp (2 and 3); The CC genotype shows one band of 324 bp (5).
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Figure 1: Restriction fragment length polymorphism analysis for determination of genotype. A. For SNP1, the GG genotype shows two bands of 362 bp and 220 bp (1 and 2); The AG genotype shows three bands of 362 bp, 258 bp and 220 bp (3, 4, 6 and 7), The AA genotype shows three bands of 258 bp, 220 bp and 104bp (5). B. For SNP2, the TT genotype shows one band of 416 bp (1 and 4); The CT genotype shows two bands of 416 bp and 324 bp (2 and 3); The CC genotype shows one band of 324 bp (5).

Mentions: Using Haploview 4.2 software and International HapMap Project website phase I &II data base (http://www.hapmap.org), we obtained two tag SNPs: SNP1 (rs605143) and SNP2 (rs565470) by using minor allele frequency (MAF) ≥ 0.05 and linkage disequilibrium patterns with r2 ≥ 0.8 as a cutoff. Genotyping in this present case–control study was confirmed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis. Sequence information for use as a reference template was obtained from the Ensembl Genome Browser (Human, numberENSG00000187498). Sequencing primers were designed using Primer Premier 5.0 software, synthesis of the Premier was undertaken by Shanghai Genery Biological Technology Company Limited (Shanghai, China). PCR amplification was performed using 25 uL of 2*powder Taq PCR master mix (Beijing Biotech, Beijing, China), 50 ng of genomic DNA, 21 uL of distilled water, 1uL of each forward and reverse primer in a 50 μL final reaction volume. The thermal cycling conditions were as follows: an initial denaturation step at 95°C for 5 min; 30 cycles of 95°C for 30s, 60°C for 35 s and 72°C for 1 min was followed by a final extension step of 72°C for 10 min. Thermal cycling was performed using the GeneAmp 9700 system (Applied Biosystems) and. PCR products were digested by restriction enzyme (Fermentas, Beijing, China) in a 20 μL final reaction volume, along with 10 μL of PCR product, 5U of restriction enzyme, 9 uL of distilled water and 1uL Solution Buffer, incubated overnight at 37°C. The primer pair sequences, annealing temperatures, resulting fragments and restriction enzymes for the two SNPs are detailed in Table 1. Resulting fragments were separated on 3.0% agarose gel (Figure 1). Finally to ensure the results to be verified, we used sequenced genomic DNAs as positive controls in our assays.


Association of COL4A1 genetic polymorphisms with coronary artery disease in Uygur population in Xinjiang, China.

Adi D, Xie X, Ma YT, Fu ZY, Yang YN, Li XM, Xiang Y, Liu F, Chen BD - Lipids Health Dis (2013)

Restriction fragment length polymorphism analysis for determination of genotype. A. For SNP1, the GG genotype shows two bands of 362 bp and 220 bp (1 and 2); The AG genotype shows three bands of 362 bp, 258 bp and 220 bp (3, 4, 6 and 7), The AA genotype shows three bands of 258 bp, 220 bp and 104bp (5). B. For SNP2, the TT genotype shows one band of 416 bp (1 and 4); The CT genotype shows two bands of 416 bp and 324 bp (2 and 3); The CC genotype shows one band of 324 bp (5).
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4016473&req=5

Figure 1: Restriction fragment length polymorphism analysis for determination of genotype. A. For SNP1, the GG genotype shows two bands of 362 bp and 220 bp (1 and 2); The AG genotype shows three bands of 362 bp, 258 bp and 220 bp (3, 4, 6 and 7), The AA genotype shows three bands of 258 bp, 220 bp and 104bp (5). B. For SNP2, the TT genotype shows one band of 416 bp (1 and 4); The CT genotype shows two bands of 416 bp and 324 bp (2 and 3); The CC genotype shows one band of 324 bp (5).
Mentions: Using Haploview 4.2 software and International HapMap Project website phase I &II data base (http://www.hapmap.org), we obtained two tag SNPs: SNP1 (rs605143) and SNP2 (rs565470) by using minor allele frequency (MAF) ≥ 0.05 and linkage disequilibrium patterns with r2 ≥ 0.8 as a cutoff. Genotyping in this present case–control study was confirmed by polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) analysis. Sequence information for use as a reference template was obtained from the Ensembl Genome Browser (Human, numberENSG00000187498). Sequencing primers were designed using Primer Premier 5.0 software, synthesis of the Premier was undertaken by Shanghai Genery Biological Technology Company Limited (Shanghai, China). PCR amplification was performed using 25 uL of 2*powder Taq PCR master mix (Beijing Biotech, Beijing, China), 50 ng of genomic DNA, 21 uL of distilled water, 1uL of each forward and reverse primer in a 50 μL final reaction volume. The thermal cycling conditions were as follows: an initial denaturation step at 95°C for 5 min; 30 cycles of 95°C for 30s, 60°C for 35 s and 72°C for 1 min was followed by a final extension step of 72°C for 10 min. Thermal cycling was performed using the GeneAmp 9700 system (Applied Biosystems) and. PCR products were digested by restriction enzyme (Fermentas, Beijing, China) in a 20 μL final reaction volume, along with 10 μL of PCR product, 5U of restriction enzyme, 9 uL of distilled water and 1uL Solution Buffer, incubated overnight at 37°C. The primer pair sequences, annealing temperatures, resulting fragments and restriction enzymes for the two SNPs are detailed in Table 1. Resulting fragments were separated on 3.0% agarose gel (Figure 1). Finally to ensure the results to be verified, we used sequenced genomic DNAs as positive controls in our assays.

Bottom Line: For total and men, the rs605143 was found to be associated with CAD by in a dominate model (p = 0.014, p = 0.013, respectively).The rs565470 was also found to be associated with CAD in a recessive model for total and men (both p < 0.001), and the difference remained statistically significant after multivariate adjustment (P = 0.002, P = 0.001, respectively).Both rs605143 and rs565470 of COL4A1gene are associated with CAD in Uygur population of China.

View Article: PubMed Central - HTML - PubMed

Affiliation: Department of Cardiology, First Affiliated Hospital of Xinjiang Medical University, Urumqi 830054, People's Republic of China. myt_xj@sina.com.

ABSTRACT

Background: Type IV collagen is important for the structural integrity and function of basement membranes. Basement membranes surround vascular smooth muscle cells in the media, COL4A1 is the most abundant component of type IV collagen in all Basement membranes. However, the relationship between COL4A1 genetic polymorphisms and coronary artery disease (CAD) remains unclear. We performed a case-control study to explore the association of COL4A1 genetic polymorphisms with CAD in Uygur population of China.

Methods: 1095 Uygur people (727 men, 368 women) including 471 CAD patients and 624 controls were selected for the present study. Two SNPs (rs605143 and rs565470) were genotyped by using the polymerase chain reaction-restriction fragment length (PCR-RFLP) method.

Results: For total and men, the rs605143 was found to be associated with CAD by in a dominate model (p = 0.014, p = 0.013, respectively). The difference remained statistically significant after multivariate adjustment (p = 0.036, p = 0.014, respectively). The rs565470 was also found to be associated with CAD in a recessive model for total and men (both p < 0.001), and the difference remained statistically significant after multivariate adjustment (P = 0.002, P = 0.001, respectively).

Conclusion: Both rs605143 and rs565470 of COL4A1gene are associated with CAD in Uygur population of China.

Show MeSH
Related in: MedlinePlus