Microsatellite alteration and metylation of RASSF1 gene in renal cell carcinoma

  • H. V. Dubrovska
  • K. V. Onyshchenko
  • L. V. Pereta
  • O. V. Kashparova
  • V. M. Grygorenko
  • I. Ya. Skrypkina


Aim. It becomes more and more relevant to study as the promising diagnostic and prognostic markers the epigenetic and genetic changes that determine the malignant transformation of cells and biological behavior of tumors. The purpose of our research was to study the loss of heterozygosity of the RASSF1 gene and the status of methylation of the locus of the RASSF1A gene in malignant tumors of the kidneys and to assess the potential for its use as a marker in clinical practice. Methods. The determination of the allele imbalance in the RASSF1 gene locus was performed by PCR using highly polymorphic markers D3S966, D3S1568 for renal cancer (RC) in paired samples of tumor and conditionally normal tissue and the PCR products detection in polyacrylamide gels. The epigenetic variability analysis of the RASSF1A gene was performed by using methyl-specific PCR, which was preceded by bisulfite DNA treatment. The statistical significance of the differences between the study groups was analyzed by using F-test and the U-test. Results. Our studies showed the predictive value of inactivation of RASSF1A in RC. The methylation level of the genomic DNA of the RASSF1A loci of the RASSF gene was 72 %. The loss of the heterozygosity of the RASSF1 gene in this type of oncological disease was about 35.4 %. Conclusions. The results may indicate that the gene RASSF1can become a candidate for inclusion in the prognostic system of clinical course of the disease.

Keywords: kidney cancer, epigenetic changes, methylation, loss of heterozygosity, RASSF1.


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