Genotyping the –6A/G functional polymorphism in the core promoter region of angiotensinogen gene by microchip electrophoresis

Angiotensinogen (AGT) gene has been regarded as one of the candidate genes for essential hypertension. In our study, the role of AGT gene as a putatively predisposing gene for hypertension was evaluated by genotyping a A (‐6) G polymorphism in the core promoter region in 123 patients with essential hypertension and 103 healthy controls. A microchip electrophoresis method coupled with polymorphism chain reaction (PCR)‐restriction fragment length polymorphism (RFLP) assay was used for genotyping the A (‐6) G single‐nucleotide polymorphism. The separation and detection of the digested PCR amplicons were completed just in 280 s or less. The genotype frequency fulfilled the criteria of the Hardy‐Weinbery equilibrium (X2 = 3.067, P > 0.05). The results showed a higher frequency of the –6 A allele (0.70) in the normotensive subjects, which is higher than those reported in Germany (0.47) and Czech (0.40) populations, but similar to that found in Japanese populations (0.73). The frequencies of genotype AA, AG, and GG were 0.46, 0.49, and 0.05 in hypertensive subjects, and 0.44, 0.53, and 0.03 in control subjects. There is no significant difference in the distributions of the genotype and allele between the two groups (X2 = 0.88, P > 0.05; X2 = 0.024, P > 0.05). These findings differ from some of the results obtained in other ethnic groups, indicating the potential importance of ethnic origin in the assessment of genetic risk identifiers for a complex disease.

[1]  A. Onipinla,et al.  Relationship between the M235T and G(-6)A polymorphisms of the angiotensinogen gene , 1999, Journal of Human Hypertension.

[2]  J. Redón,et al.  Polymorphisms of the angiotensinogen gene and the outcome of microalbuminuria in essential hypertension: a 3-year follow-up study , 2004, Journal of Human Hypertension.

[3]  C. Yunis,et al.  Association of angiotensinogen M235T and A(-6)G gene polymorphisms with coronary heart disease with independence of essential hypertension: the PROCAGENE study. Prospective Cardiac Gene. , 2001, Journal of the American College of Cardiology.

[4]  G. Shan,et al.  A-6G variant of the angiotensinogen gene and essential hypertension in Han, Tibetan, and Yi populations. , 2001, Hypertension research : official journal of the Japanese Society of Hypertension.

[5]  W. März,et al.  Angiotensinogen M235T polymorphism is associated with plasma angiotensinogen and cardiovascular disease. , 1999, American heart journal.

[6]  R. McCormick,et al.  Microchannel electrophoretic separations of DNA in injection-molded plastic substrates. , 1997, Analytical chemistry.

[7]  John Quackenbush,et al.  A nucleotide substitution in the promoter of human angiotensinogen is associated with essential hypertension and affects basal transcription in vitro. , 1997, The Journal of clinical investigation.

[8]  D. Pop,et al.  Essential arterial hypertension and polymorphism of angiotensiongen M235T gene , 2002, Journal of cellular and molecular medicine.

[9]  T. Yee,et al.  Association of angiotensinogen gene T235 variant with increased risk of coronary heart disease , 1995, The Lancet.

[10]  D. Ilsley,et al.  A microfluidic system for high‐speed reproducible DNA sizing and quantitation , 2000, Electrophoresis.

[11]  R. Schmieder,et al.  Angiotensin II related to sodium excretion modulates left ventricular structure in human essential hypertension. , 1996, Circulation.

[12]  J. Lalouel,et al.  Haplotypes of angiotensinogen in essential hypertension. , 1997, American journal of human genetics.

[13]  L. Mosso,et al.  A(-6)G variant of angiotensinogen gene and aldosterone levels in hypertensives. , 1999, Hypertension.

[14]  P. Corvol,et al.  Structure of human angiotensinogen gene. , 1989, DNA.

[15]  B. Morris,et al.  Exclusion of angiotensinogen gene in molecular basis of human hypertension: sibpair linkage and association analyses in Australian anglo-caucasians. , 1999, American journal of medical genetics.

[16]  M. Peach,et al.  Molecular Biology of Angiotensinogen , 1991, Hypertension.

[17]  B. Lin,et al.  DNA diagnosis by capillary electrophoresis and microfabricated electrophoretic devices , 2003, Expert review of molecular diagnostics.

[18]  Y. Toya,et al.  Angiotensinogen gene polymorphism near transcription start site and blood pressure: role of a T-to-C transition at intron I. , 1999, Hypertension.

[19]  Y. Nibu,et al.  A cis-Acting DNA Element Located between TATA Box and Transcription Initiation Site Is Critical in Response to Regulatory Sequences in Human Angiotensinogen Gene* , 1996, The Journal of Biological Chemistry.

[20]  R. Schmieder,et al.  Angiotensinogen Gene Core Promoter Variants and Non-Modulating Hypertension , 2001, Hypertension.

[21]  P Corvol,et al.  Molecular genetics of human hypertension: role of angiotensinogen. , 1997, Endocrine reviews.

[22]  P. Munroe,et al.  Linkage of the angiotensinogen gene to essential hypertension. , 1994, The New England journal of medicine.

[23]  T. Ogihara,et al.  Nine polymorphisms of angiotensinogen gene in the susceptibility to essential hypertension. , 2000, Life sciences.

[24]  A. Vasku,et al.  An association of BMI with A (-6) G, M235T and T174M polymorphisms in angiotensinogen gene in essential hypertension , 2002, Journal of Human Hypertension.

[25]  R. Felder,et al.  Combinations of variations in multiple genes are associated with hypertension. , 2000, Hypertension.