Search

Persistent infection with high-risk types of human papillomaviruses (HPV) is a major cause of cervical cancer, and an important factor in other malignancies, for example, head and neck cancer. Despite recent progress in screening and vaccination, the incidence and mortality are still relatively high, especially in low-income countries. The mortality and financial burden associated with the treatment could be decreased if a simple, rapid, and inexpensive technology for HPV testing becomes available, targeting individuals for further monitoring with increased risk of developing cancer. Commercial HPV tests available in the market are often relatively expensive, time-consuming, and require sophisticated instrumentation, which limits their more widespread utilization. To address these challenges, novel technologies are being implemented also for HPV diagnostics that include for example, isothermal amplification techniques, lateral flow assays, CRISPR-Cas-based systems, as well as microfluidics, paperfluidics and lab-on-a-chip devices, ideal for point-of-care testing in decentralized settings. In this review, we first evaluate current commercial HPV tests, followed by a description of advanced technologies, explanation of their principles, critical evaluation of their strengths and weaknesses, and suggestions for their possible implementation into medical diagnostics., (© 2024 The Authors. Journal of Medical Virology published by Wiley Periodicals LLC.)

Cervical cancer is one of the most common malignancies in women. MicroRNAs (miRNAs) are involved in a variety of fundamental cellular processes, including carcinogenesis. The potential utilization of aberrantly expressed miRNAs as novel biomarkers in cervical cancer diagnostics is growing. We investigated miRNA expression profiles during the progression of dysplasia in cervical epithelium to identify aberrantly expressed miRNAs. High-throughput miRNA profiling of high-grade precancerous lesions identified 79 miRNAs showing significant difference in expression values compared to normal cervical epithelium. Ten selected miRNAs were subsequently measured in an independent group of samples to validate them as promising biomarkers of cervical carcinogenesis. MicroRNAs miR-10b-5p, miR-34c-5p, miR-409-3p and miR-411-5p were confirmed as downregulated, while miR-10a-5p, miR-132-3p, miR-141-5p were significantly upregulated in dysplastic cervical tissues. Further investigation revealed an inverse correlation of miR-409-3p with E6 mRNA levels in precancerous cervical lesions. Subsequent in vitro analyses showed a direct involvement of this miRNA in the regulation of E6 oncogene levels, thus confirming a potential tumor suppressor function of miR-409-3p in cervical malignancies. Hence, miR-409-3p may represent a useful early marker and a potential therapeutic target for cervical cancer., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: Cervical cancer is the fourth most common cancer in women and is usually associated with human papillomavirus infection. Viral infections are usually characterized by morphological changes of epithelial cells; however, it is difficult to determine using recently available screening methods whether the changes are caused by productive infection or by malignant disease. Thus, new efforts are required to find novel diagnostic biomarkers of cervical cancer, such as miRNAs, which are small non-coding RNAs involved in the regulation of gene expression., Materials and Methods: miR-34c levels in cervical cancer cell lines were determined by the droplet-digital polymerase chain reaction. Changes in miR-34c expression in vitro were achieved by transient transfection with a specific miRNA mimic and inhibitor oligonucleotides. Cell proliferation was analyzed by crystal violet staining followed by spectrophotometric measurements. The effect on migratory properties was studied using a „scratch“ assay. Western blotting analysis was used to determine the expression of selected proteins., Results: The downregulation of miR-34c expression was associated with a slight increase in cellular proliferation and a significant increase in cell migration. The analysis of miR-34c expression performed on a set of 39 dysplastic tissues and 35 samples of healthy controls subsequently revealed a significant difference (p < 0.01) in the level of this miRNA., Conclusion: Comparative expression analysis revealed lower expression of miR-34c in cervical precanceroses than in normal untransformed epithelium. in vitro modulation of miR-34c expression revealed its tumor suppressor role in cervical  malignancies. Key words: cervical cancer - HPV - miRNA - HSIL - hsa-miR-34c - precancerosis This work was supported by the projects MEYS - NPS I - LO1413, P206/12/G151 and MH CZ - DRO (MMCI, 00209805). The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study.  The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Accepted: 16. 7. 2018.

Background: Persistent infection with high-risk human papillomavirus (HPV) strains, especially HPV 16 and HPV 18, is associated with the onset of various malignant diseases, including cervical carcinoma in women. HPV DNA testing is thus being implemented as a complementary method to standard cytological examination, mainly due to its increased sensitivity., Aim: This review outlines the role of HPV in cervical carcinogenesis, with a focus on the formation of cervical intraepithelial neoplasias (CIN1-3) and the molecular mechanism underlying cellular transformation. Current biomarkers used to screen premalignant lesions are described, including mRNA transcripts of the E6 and E7 genes, protein p16 (a cyclin-dependent kinase inhibitor that regulates cell cycle progression from G1 to S phase), altered DNA methylation patterns, and actions of specific microRNAs (short (18-22 bp), non-coding, single-stranded RNA molecules that regulate gene expression at the post-transcriptional level). This review also describes the advantages and drawbacks of commercial HPV tests, and depicts novel methods for more cost-effective and faster HPV diagnostics based on optical or electrochemical detection., Conclusion: Although great progress has been made, the incidence and mortality rates of cervical malignancies remain relatively high, especially in developing countries. Incorporation of HPV testing into routine screening programs could help to decrease mortality rates; however, the cost of such testing must be reduced if it is to compete with current cytology-based examinations.Key words: HPV - cervical carcinoma - HPV testing - nucleic acid hybridization - mRNA - DNA methylation - microRNA This work was supported by MEYS-NPS I-LO1413 and GAČR 17-08971S. The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers.Submitted: 25. 9. 2017Accepted: 26. 1. 2018.

Cervical cancer, being the fourth leading cause of cancer death in women worldwide, predominantly originates from a persistent infection with a high-risk human papillomavirus (HPV). Detection of DNA sequences from these high-risk strains, mostly HPV-16 and HPV-18, represents promising strategy for early screening, which would help to identify women with higher risk of cervical cancer. In developing countries, inadequate screening options lead to disproportionately high mortality rates, making a fast and inexpensive detection schemes highly important. Electrochemical sensors and assays offer an alternative to current methods of detection. We developed an electrochemical-chip based assay, in which target HPV DNA is captured via magnetic bead-modified DNA probes, followed by an antidigoxigenin-peroxidase detection system at screen-printed carbon electrode chips, enabling parallel measurements of eight samples simultaneously. We show sensitive detection in attomoles of HPV DNA, selective discrimination between HPV-16 and HPV-18 and good reproducibility. Most importantly, we show application of the assay into both cancer cell lines and cervical smears from patients. The electrochemical results correlated well with standard methods, making this assay potentially applicable in clinical practice., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Introduction: Human papillomavirus (HPV) infection represents the most important risk factor for the development of cervical intraepithelial neoplasia (CIN) and cervical cancer. We aimed to analyze the consequences of methylation of the E6 gene promoter in distinct stages of HPV-16-induced cellular transformation to assess its importance for disease progression., Methods: Human papillomavirus 16 was detected by sensitive polymerase chain reaction (PCR). Determination of E6 gene promoter methylation was analyzed by digestion with specific restriction endonuclease McrBC followed by PCR amplification. Expression of the E6 gene was determined by quantitative real-time PCR., Results: Of 103 cervical smears from asymptomatic women with no cytological and colposcopic abnormalities, 20.4% were HPV-16-positive. Human papillomavirus 16 was present in 44.4% of 18 patients with CIN I, in 62.2% of 143 patients with CIN II/III, and in 74.2% of 31 cervix carcinoma specimens. The incidence of HPV-16 in all lesions compared with asymptomatic women was statistically significant (P < 0.001, Pearson chi test). Methylation was detected in 81% (n = 21) of HPV-16-positive asymptomatic smears compared with 62.5% in CIN I (n = 8), 31.5% (n = 89) in CIN II/III, and 43.4% (n = 23) in carcinomas; a statistical significance between lesions and healthy women was found (P < 0.001, Pearson chi test). Expression of E6 mRNA correlated with methylation status (P = 0.010, Mann-Whitney U test)., Conclusions: We conclude that methylation of the E6 gene promoter in HPV-16 genome is a predictive biomarker for cervical cancer progression by regulating the expression of the E6 oncogene.

Objective: To summarize the significance of methylation in HPV16 genome to cervix cancerogenesis., Design: Review., Setting: Department of Oncological and Experimental Pathology, Masaryk Memorial Cancer Institute, Brno., Subject and Method: Human papillomaviruses, especially HPV16, are the most frequent causative agents of cervical intraepithelial neoplasia and cervical carcinoma. Their ability to initiate transformation of infected epithelial cells fully depends up production of viral early phase proteins E6 and E7. Affected keratinocytes activate defensive mechanisms based on inhibition of viral DNA transcription by changes in chromatin structure like DNA methylation or histon deacetylation and therefore prevent transcriptional factors from binding to target promoters and from the production of viral oncoproteins., Conclusion: Research into epigenetic mechanisms of gene silencing clearly showed their important roles in etiology of cancer. Recent findings confirm the significance of methylation of HPV16 oncogenes leading to block of neoplastic transformation, and simultaneously they indicate new therapeutic possibilities linked with reactivation of methylated tumor supressors.