Alteration of the exDNA profile in blood serum of LLC-bearing mice under the decrease of tumour invasion potential by bovine pancreatic DNase I treatment

PLoS One. 2017 Feb 21;12(2):e0171988. doi: 10.1371/journal.pone.0171988. eCollection 2017.

Abstract

Taking into account recently obtained data indicating the participation of circulating extracellular DNA (exDNA) in tumorigenesis, enzymes with deoxyribonucleic activity have again been considered as potential antitumour and antimetastatic drugs. Previously, using murine Lewis lung carcinoma and hepatocellular carcinoma A1 tumour models, we have shown the antimetastatic activity of bovine DNase I, which correlates with an increase of DNase activity and a decrease of exDNA concentration in the blood serum of tumour-bearing mice. In this work, using next-generation sequencing on the ABS SOLiD™ 5.500 platform, we performed a search for molecular targets of DNase I by comparing the exDNA profiles of healthy animals, untreated animals with Lewis lung carcinoma (LLC) and those with LLC treated with DNase I. We found that upon DNase I treatment of LLC-bearing mice, together with inhibition of metastasis, a number of strong alterations in the patterns of exDNA were observed. The major differences in exDNA profiles between groups were: i) the level of GC-poor sequences increased during tumour development was reduced to that of healthy mice; ii) levels of sequences corresponding to tumour-associated genes Hmga2, Myc and Jun were reduced in the DNase I-treated group in comparison with non-treated mice; iii) 224 types of tandem repeat over-presented in untreated LLC-bearing mice were significantly reduced after DNase I treatment. The most important result obtained in the work is that DNase I decreased the level of B-subfamily repeats having homology to human ALU repeats, known as markers of carcinogenesis, to the level of healthy animals. Thus, the obtained data lead us to suppose that circulating exDNA plays a role in tumour dissemination, and alteration of multiple molecular targets in the bloodstream by DNase I reduces the invasive potential of tumours.

MeSH terms

  • Animals
  • Carcinoma, Lewis Lung / blood*
  • Carcinoma, Lewis Lung / pathology
  • Cattle
  • DNA, Neoplasm / blood*
  • DNA, Neoplasm / genetics
  • Deoxyribonuclease I / metabolism*
  • Extracellular Space / chemistry
  • Gene Library
  • High-Throughput Nucleotide Sequencing
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Neoplasm Invasiveness*
  • Neoplasm Transplantation
  • Real-Time Polymerase Chain Reaction

Substances

  • DNA, Neoplasm
  • Deoxyribonuclease I

Grant support

This research was supported by the Russian Academy of Science under the programs 1.7 ‘Molecular and Cellular Biology’ (grant # II.2P/VII.58-1), Russian Foundation for Basic Research (grant # 14-04-01007) and scientific schools SS - 7623.2016.4. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.