Complex analysis of p53-responsive microRNA genes methylation and TР53 gene mutations in Diffuse Large B-cell Lymphoma

Relevance. Comparison of microRNA expression in lymphoma and normal lymphoid tissue revealed a decrease in the expression level of a number of p53-induced oncosuppressive molecules, such as miR-34a, miR-34b/c, miR-129 and miR-203, which, in addition to mutations in the TP53 gene, can be caused by aberrations in the genome regions encoding the microRNAs themselves. Purpose. To carry out a comprehensive analysis of the p53-responsive microRNAs genes MIR-34A, MIR-34B/C, MIR-203 and MIR-129-2 methylation and mutations of the TP53 gene in diffuse large B-cell lymphoma (DLBCL). Methods. 73 samples of DNA isolated from tumor tissue of patients with DLBCL were analyzed. The nucleotide sequence of the TP53 gene DNA-binding domain was determined by capillary direct Sanger sequencing. The methylation status was determined by methyl-specific PCR (for MIR-203 and MIR-129-2) and methyl-sensitive analysis of high-resolution melting curves (for MIR-34A and MIR-34B/C) using DNA treated with sodium bisulfite. Results. The frequency of MIR-34A, MIR-34B/C, MIR-203 and MIR-129-2 genes methylation and TP53 gene mutations was 27%, 62%, 66%, 67% and 25%, respectively. Methylation of the analyzed genes of p53-responsive microRNAs and mutations in the TP53 gene in the tumor tissue of DLBCL in the most patients tended to mutual exclusion. A significant association (p < 0.05) was shown between the methylation of the MIR-203, MIR-129-2 and MIR-34B/C genes, as well as the MIR-34B/C and MIR-34A pair. In 14% and 47% of DLBCL cases all four genes and three of the analyzed genes were methylated, respectively. Conclusions. Along with mutations in the TP53 gene aberrant methylation may be an independent cause of decreased miR-34a, miR-34b/c, miR-129 and miR-203 expression. Methylation of the MIR-34A, MIR-34B/C, MIR-203 and MIR-129-2 genes in the tumor tissue of DLBCL in most cases is of a combined nature.

ТР53 gene, microRNA, methylation, miR-34a, miR-34b/c, miR-129, miR-203, lymphoma

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