Kharchenko A. V.

CLINICAL AND EXPERIMENTAL MEDICINE

Scentific article

Diagnosis, made by the ISSR-PCR reaction, has shown mucosal epithelial DNA changes specific to the epithelial dysplasia of varied severity in gastric mucosa in patients with chronic duodenal ulcer. Amplicated profiles of normal gastric mucosa marker enclosed fragments measuring 190 – 60 p. n. (pairs of nucleotides) and were identical within its group and significantly differed from the DNA-profiles of other subject groups. The DNA-profiles measured 220 – 90 p. n. corresponded to phenotype of Stage I epithelial dysplasia (D-І). Stage II epithelial dysplasia (D-IІ) phenotypes in 90 % of cases possessed the variant of DNA-profiles measured 300 – 100 p. n. (the first variant) and the rest ones had the DNA-profiles measured 500 – 100 p. n. (the second variant). Stage III epithelial dysplasia (D-IІI) phenotypes had the DNA-profiles of single variant, measured 520 – 320 p. n. D-IІ phenotype corresponded to two variants of DNA-profiles, containing amplicones measured within 500 p. n. and without them. The latter in 36,4 % resembled DNA-profiles of Type III gastric mucosa. It makes evident that D-II DNA-profiles are changing and have transitional forms to D-III DNA-profiles. In cases with abovementioned dysplasia, changes in the form of enlargement of amplicones, specific to each group, have been observed. These changes are characterized as microsatellite expansion. Genetic typing of gastric mucosa epithelium, made by the ISSR – PCR reaction found that gastric mucosa epi- thelial dysplasias in patients with chronic duodenal ulcer had undergone specific changes, related to their corre- spondent DNA-profiles. Amplificated normal DNA-profile has the spectrum of amplicones measured within 190 – 60 p. n. ; low-grade dysplasia (D-I) has amplificated DNA-profile with spectrum of amplicones measured from 220 to 60 p. n. ; medium- grade dysplasia (D-II) in 90 % has amplificated DNA-profile with spectrum of amplicones measured 300 – 100 p. n. (I – variant) and in 10 % has DNA-profile of 500 – 100 p. n. (ІІ – variant), indicating about heterogeneity of Stage II dysplasias. D-III dysplasia has single variant of amplificated DNA-profile with spectrum of amplicones measured 520 – 320 p. n. The ISSR – PCR reaction made extension of microsatellite DNA-sequences evident. That is, the results of genetic typing of gastric mucosa epithelium founded microsatellite expansions. It is known that that inten- sive extension of microsatellite sequences due to replicative errors is called microsatellite expansions. There is strong correlation between the stage of gastric mucosa epithelial dysplasia on phenotypical character- istics and indices of DNA-typing of gastric mucosa samples; Pearson correlation coefficient rxy was 0,863 and 0,917, respectively. The total results founded the existences of statistically significant dependence with 0,99 probability. The ISSR – PCR reaction is the informative method to detect change of gastric mucosa epithelium genetic structure. Taking into account the availability, relative simplicity and possibility of visual reading of the results without special equipment it has been successfully used while studying the DNA-typing of gastric mucosa epithelium with phenotypes of epithelial dysplasia, providing with detection of change, they are experiencing.

DNA, amplicones, phenotype

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«Bulletin of problems biology and medicine» Issue 1 (106), 2014 year, 199-203 pages, index UDK 616 – 07: 616. 33 – 008. 3