Анализ аберрантно-метилированных последовательностей в составе циркулирующих ДНК крови при раке легкого

Abstract

Malignant cell transformation is accompanied by two processes of DNA methylation changes: promoter hypermethylation of specific genes and hypomethylation of retrotransposons. The composition of circulating DNA (cirDNA) from plasma and cell-surface-bound circulating DNA (csb-cirDNA) was shown earlier to be altered in the blood of cancer patients due to accumulation of tumor-specific aberrantly methylated DNA fragments, which are currently considered valuable cancer markers. The present study compares LINE-1 retrotransposon methylation patterns in plasma cirDNA and csb-cirDNA from untreated lung cancer patients (LC) and healthy donors. Concentrations of methylated LINE-1 region 1 copies (LINE-1met) were assayed by real-time methylation-specific PCR. In order to normalize the LINE-1 methylation level, the LINE-1 region 2 concentration was evaluated, which was independent of the methylation status (LINE-1Ind). We recorded an statistically significant increase of the LINE-1 methylation index determined as (LINE-1met/LINE-1Ind) due to the profound LINE-1Ind decrease (Mann-Whitney test, p = 0.005). Plasma cirDNA demonstrated no difference in the ratio LINE-1met/LINE-1Ind between LC patients and healthy donors (p = 0.40). The data obtained agree with our earlier results, which showed that csb-cirDNA was a highly informative material for lung cancer diagnostics.