Simultaneous Detection of the HPV L1 Gene and the Human β-Globin Gene in the Blood Components of Cervical Cancer Patients Living in Yakutia

Abstract

Background: To create a test for the early detection of cervical cancer (CC), we conducted exploratory studies on detecting HPV genes and genes of the human β-globin locus in plasma (Pl) and RBC suspension (RBCsus) samples from patients with newly detected CC (NDCC). Methods and Results: Smears of venous blood containing K3-EDTA from five anonymous patients aged from 45 to 55 years (residents of Yakutia), with NDCC were obtained. Three types of blood component samples were prepared – plasma (Pl), red blood cell (RBC) suspension (RBCsus), and the erythrocyte fraction treated with trypsin (RBCsus-Try). To detect circulating cell-free DNA (cfDNA) in NDCC patients, we studied the presence of genes corresponding to the HPV L1 protein region and genes of the human β-globin locus by real-time PCR (qPCR) using appropriate primers. The genes of β-globin locus and HPV L1 were detected in Pl of NDCC patients in 20% of cases, and in RBCsus in 60% of cases. The amplified gene products using primers were present in RBCsus-Try in only one patient (20% of cases). In patients with uncertain amplification, electrophoresis showed the absence of amplified products in Pl and their presence in RBCsus. Conclusion: In NDCC patients, HPV L1 and β-globin genes can be detected in both Pl and RBCsus. In addition, in RBC samples, these genes were detected more often than in plasma samples, and no absolute absence of amplification products was observed in RBC samples. The research needs to be continued.

Blood Plasma Exosomes Contain Circulating DNA in Their Crown

“…However, at present, there is still no unequivocal opinion regarding the sources of cirDNA, the mechanisms that ensure their circulation and elimination from the bloodstream, and, most importantly, the biological role of cirDNA [13][14][15]. It is known that cirDNA is resistant to the action of extracellular nucleases due to the formation of macromolecular complexes with proteins (including histones) [16] and other biomolecules, as well as the binding of these complexes to the surface of blood cells [17,18]. Notably, several pathological conditions, including cancer and autoimmune diseases, which are characterized by increased cirDNA concentrations [19,20], are also characterized by elevated exosome levels in plasma or serum [21].…”

The Influence of Proteins on Fate and Biological Role of Circulating DNA

“…Moreover, leukocytes were shown not only to carry DNA on their surface, but to also bind it by the ligand-receptor mechanism [34]. Later, the presence of cirDNA, eluted by mild trypsin treatment of the erythrocyte, leukocyte, and platelet surfaces, was confirmed [34,62]. Similar high molecular weight cirDNA fragments were also detected on the cell surface in vitro [27].…”