Variability of some mitochondrial dna point mutations in buccal and whole blood human cells

Abstract

s / Atherosclerosis 235 (2014) e27–e83 e65 51 Novel risk factors and biomarkers EAS-1065. VARIABILITY OF SOME MITOCHONDRIAL DNA POINT MUTATIONS IN BUCCAL AND WHOLE BLOOD HUMAN CELLS V.V. Sinyov , M.A. Sazonova , S.A. Kosogorova , Y.V. Bobryshev , A.Y. Postnov , A.N. Orekhov , I.A. Sobenin a Cardiovascular Pathology, Cardiology Research Complex MH RF, Moscow, Russia; b Laboratory of Mechanisms Atherogenesis, Institute of General Pathology and Patophysiology RAMS, Moscow, Russia; c Laboratory of Medical Genetics, Institute for Atherosclerosis Research Skolkovo Innovative Centre, Moscow, Russia Objectives: According to the literature data, single nucleotide polymorphisms of mitochondrial genome are unevenly widespread in organs and tissues. This prohibits from the usage of such biomarkers in diagnosing various diseases. That is why a search of biomarkers for non-invasive genetic diagnostics seems to be extremely urgent. The aim of the present study was an assessment of capability for the buccal epithelium usage instead of blood leukocytes in genetic diagnostics of liability to atherosclerosis. A pilot study of heteroplasmy level in mitochondrial genome mutations G15059A, G13513A, G12315A, C3256T and A1555G in human buccal epithelium and whole blood was carried out. An association of these mutations with atherosclerosis had been previously found while analyzing blood leukocytes is a sample of 700 patients, examined in the Moscow Region. Methods: Buccal epithelium and whole blood samples of 48 donors were used as a material for investigation. A total DNA purification from tissue samples was performed by the method of phenolchloroform extraction. Afterwards the assayed samples of mDNA were amplified with PCR and analyzed by new original method developed in our laboratory on the basis of pyrosequence technology. The investigations were carried out with the help of an automatic pyrosequenator PSQ96? A.The statistical evaluation of the results was performed by using SPSS version 22.0. Results: Significant differences of heteroplasmy level (p0,05) were found between mitochondrial genome mutations G13513A and C3256T in buccal epithelium and blood cells. Meanwhile in mutations G15059A, G12315A and A1555G such differences are absent. Conclusion: According to the obtained data, mitochondrial genome mutations G15059A (cytochrome B gene), G12315A (tRNA-Leucine gene, codon recognized CUN) and A1555G (rRNA 12S gene) can be used in buccal epithelium analysis for assessment of the liability to atherosclerosis. This study was supported by the Russian Ministry of Education and Science.