Matrix Metalloproteinase-9 Gene Polymorphism and Its Methylation in Stroke Patients

Abstract

Background: Genetic and environmental factors, along with hypertension, diabetes mellitus and smoking cause accelerated atherosclerosis and, eventually, stroke. Matrix metalloproteinase-9 (MMP-9) are inflammatory mediators of the endoproteinase family, and their polymorphism and methylation are associated with the development of atherosclerosis and stroke. This study explores this association in the Indian population.

Objective: To study the association of MMP gene polymorphism and methylation with the risk of stroke.

Methods: A case-control study was conducted on 100 admitted patients (both genders) diagnosed with ischaemic stroke. Another 100 healthy subjects, not suffering from any chronic illness or stroke, were taken as controls. All participants were genotyped for rs3918242 (MMP-9) by polymerase chain reaction (PCR) and restriction fragment length polymorphism. Methylation of the MMP-9 gene-promoter region was assessed by methylation-specific PCR.

Results: The case (mean age = 61.3 ± 7.36 years old) and control (mean age = 60.68 ± 7.1 years old) groups were age-matched. Among cases, 61 patients were smokers, 55 were diabetic and 53 were hypertensive. A significant risk of ischaemic stroke was associated with the CT genotype (adjusted odds ratio [aOR] = 7.09; P < 0.001), TT genotype (aOR = 19.75; P < 0.001) and T allele (aOR = 10.71; P < 0.001). MMP-9 methylation decreased the risk of stroke (aOR = 0.23; P < 0.001).

Conclusion: MMP-9 gene-1562C/T polymorphism (SNP rs3918242) (single-nucleotide polymorphism [SNP] rs3918242) is a potential marker to predict ischaemic stroke and constitutes a significant proportion of the general population. Its polymorphism predisposes to ischaemic stroke, while its methylation is protective.

Keywords: MMP; MMP polymorphism; MMP-9 methylation; epigenetics; ischaemic; stroke.

Figure 1

Samples of MMP-9 PCR-RFLP (435 bp) CC genotype with one fragment band, TT genotype with two fragments are polymorphic and CT genotype with three fragments band are heterozygous

Figure 2

Agarose gel electrophoresis of MMP-9 gene MSP products from peripheral blood. M represents methylation specific primers; U represents unmethylated specific primers