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.
Currently, cervical cancer (CC) is one of the most common oncological diseases. In this regard, it is necessary to develop new research methods for a more detailed study of the occurrence and development of the disease at the molecular and cellular levels, as well as to improve the effectiveness of treatment and form a deeper understanding of the causes of relapses. The aim of this work was to study nanoparticles localized on the erythrocyte membrane—presumably HPV 16, 18, before and after radiation therapy, in patients with cervical cancer (CC). To study the surface of red blood cells by the SEM method, venous blood samples from 17 patients with a confirmed diagnosis of CC were prepared in thin layers evenly applied to a dry, fat-free glass slide, which was dried at room temperature. To detect nanoparticles on the surface of erythrocytes by the immunofluorescence analysis, we developed a special protocol for preparing erythrocyte masses from patients diagnosed with CC. As a result of using a new method of sample preparation for immunofluorescence assay and using SEM, the hypothesis of the viral nature of nanoparticles localized on the surface of the blood erythrocytes of patients with CC was confirmed: Particles of HPV 16 and 18 are located on the cytoplasmic membrane of erythrocytes. Studies suggest that viruses attach to the erythrocyte membrane, which seems to influence the development of CC, its recurrence, and metastasis.
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