RESUMEN
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.
Asunto(s)
Papillomavirus Humano 16/genética , Papillomavirus Humano 18/genética , MicroARNs/genética , Proteínas Oncogénicas Virales/genética , Proteínas Represoras/genética , Lesiones Intraepiteliales Escamosas/genética , Neoplasias del Cuello Uterino/genética , Línea Celular Tumoral , Regulación hacia Abajo , Femenino , Perfilación de la Expresión Génica , Regulación Neoplásica de la Expresión Génica , Humanos , Lesiones Intraepiteliales Escamosas/virología , Regulación hacia Arriba , Neoplasias del Cuello Uterino/virologíaRESUMEN
Current human papillomavirus (HPV)16 DNA testing has high sensitivity but low specificity, while mRNA testing (qualitative) improves the specificity. However, both techniques are not able to discriminate between transient and persistent infections. To overcome the disadvantages, we quantitatively detected E6 and E7 mRNAs by quantitative real-time polymerase chain reaction (qRT-PCR) in cervical brushing cells from 87 HPV16+ and 31 HPV16- patients. Our results showed that the expression levels of E6 mRNA or E7 mRNA were significantly increased in HPV16-positive cases than that in the negative cases. Furthermore, in HPV16+ cases, the expression levels of E6 mRNA were significantly increased in invasive cancer compared with high-grade squamous intraepithelial lesion (HSIL; p < 0.01), and HSIL compared with low-grade squamous intraepithelial lesion (LSIL; p < 0.01). There were no significant changes between LSIL and benign lesions. The expression levels of E7 mRNA presented no significant difference among the above-mentioned four groups. To test whether qRT-PCR can discriminate between transient and persistent infections, 57 HPV16+ patients were followed up for 1 year, and our results demonstrated that the expression levels of both E6 mRNA and E7 mRNA in the persistent infection group were significantly increased relative to the transient infection group ( p < 0.01 or 0.05). Thus, a quantitative detection of the expression levels of E6 mRNA in cervical brushing cells may not only be used as an ancillary tool to cytological diagnosis of cervical neoplasia, but may also help to determine the severity of the lesions and the triage of transient infection.
Asunto(s)
Papillomavirus Humano 16/genética , Proteínas Oncogénicas Virales/genética , Proteínas E7 de Papillomavirus/genética , Infecciones por Papillomavirus/diagnóstico , ARN Mensajero/genética , Proteínas Represoras/genética , Lesiones Intraepiteliales Escamosas de Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/diagnóstico , Adulto , Anciano , Femenino , Papillomavirus Humano 16/aislamiento & purificación , Humanos , Persona de Mediana Edad , Infecciones por Papillomavirus/virología , ARN Mensajero/análisis , Lesiones Intraepiteliales Escamosas de Cuello Uterino/virología , Neoplasias del Cuello Uterino/virología , Adulto JovenRESUMEN
The incidence of HPV-positive oropharyngeal squamous cell carcinoma (OPSCC), which is both biologically and clinically distinct from tobacco- and alcohol-related OPSCC, is dramatically increasing. The finding that individuals with HPV-positive local/regionally advanced OPSCC have a significantly better prognosis than their negative counterparts have led to efforts to de-escalate treatment in those patients to avoid serious side effects and to improve their long-term quality of life, while maintaining treatment efficacy. Identifying diagnostic tests that are able to distinguish cancers etiologically associated with HPV is thus becoming a pressing challenge for researchers. The purpose of this review is to provide an overview of the diagnostic tools presently available to evaluate HPV status in patients with OPSCC and, in particular, to discuss their strengths and weaknesses in identifying those infections that are the real driving force in the oropharyngeal carcinogenesis process.