miRNA expression profiling of hereditary breast tumors from BRCA1- and BRCA2-germline mutation carriers in Brazil.

BACKGROUND: MicroRNAs (miRNAs) are small non-coding RNAs involved in post-transcriptional gene expression regulation and have been described as key regulators of carcinogenesis. Aberrant miRNA expression has been frequently reported in sporadic breast cancers, but few studies have focused on profiling hereditary breast cancers. In this study, we aimed to identify specific miRNA signatures in hereditary breast tumors and to compare with sporadic breast cancer and normal breast tissues. METHODS: Global miRNA expression profiling using NanoString technology was performed on 43 hereditary breast tumors (15 BRCA1, 14 BRCA2, and 14 BRCAX), 23 sporadic breast tumors and 8 normal breast tissues. These normal breast tissues derived from BRCA1- and BRCA2- mutation carriers (n = 5) and non-mutation carriers (n = 3). Subsequently, we performed receiver operating characteristic (ROC) curve analyses to evaluate the diagnostic performance of differentially expressed miRNAs. Putative target genes of each miRNAs considered as potential biomarkers were identified using miRDIP platform and used for pathway enrichment analysis. RESULTS: miRNA expression analyses identified several profiles that were specific to hereditary breast cancers. A total of 25 miRNAs were found to be differentially expressed (fold change: > 2.0 and p < 0.05) and considered as potential biomarkers (area under the curve > 0.75) in hereditary breast tumors compared to normal breast tissues, with an expressive upregulation among BRCAX cases. Furthermore, bioinformatic analysis revealed that these miRNAs shared target genes involved in ErbB, FoxO, and PI3K-Akt signaling pathways. CONCLUSIONS: Our results showed that miRNA expression profiling can differentiate hereditary from sporadic breast tumors and normal breast tissues. These miRNAs were remarkably deregulated in BRCAX hereditary breast cancers. Therefore, miRNA signatures can be used as potential novel diagnostic biomarkers for the prediction of BRCA1/2- germline mutations and may be useful for future clinical management.

Risk Profile of High-grade Cervical Lesions and Cervical Cancer Considering the Combination of Cytology, HPV Genotype, and Age among Women Undergoing Colposcopy.

OBJECTIVE: The present study aims to establish a risk profile for high-grade cervical lesions and cervical cancer (CIN2 + ) in women undergoing colposcopy at the Hospital do Câncer de Barretos, through the analysis of Human Papillomavirus (HPV) infection, cervical cytology, and patient's age. METHODS: Retrospective cross-sectional study based on a computerized database of women aged ≥ 18 years old who underwent colposcopy at the Prevention Department of the Hospital do Câncer de Barretos from 2017 to 2019. RESULTS: A total of 3,411 women were included, 58.0% were positive for high-risk-HPV test, with a higher prevalence of CIN2+ for HPV16 (30.3%) and other HPV (45.0%). Cytological findings that suggest invasive cervical cancer (squamous cells or adenocarcinoma), regardless of the status of HPV test, showed 100% diagnosis of CIN2 + , while atypias that suggest high-grade lesions, HSIL and ASC-H, positive for HPV test, showed in 86 and 55.2%, respectively, diagnosis of CIN2 + . ASC-H cytological results among women aged > 40 years old and negative HPV were mainly associated with benign findings. We observed that ≤ CIN1 has a higher prevalence among older women with negative HPV, while for high-grade lesions there is an increase among young women HPV16- and/or 18-positive. In cancer diagnosis, we observed a predominance of HPV 16/18 regardless of the age group. CONCLUSION: The highest risks of precursor lesions and cervical cancer were found among women with positive HPV 16/18 tests and severe cytological atypia in population screening tests. In addition, cytological findings of ASC-H HPV negative in women > 40 years old usually represent benign findings in histological investigation.

OBJETIVO: Estabelecer um perfil de risco de lesões intraepiteliais de alto grau e câncer do colo do útero (NIC2 + ) em mulheres submetidas a colposcopia considerando-se a infecção pelo papilomavírus humano (HPV), citologia cervical e idade. MéTODOS: Estudo retrospectivo transversal em banco de dados informatizado de mulheres com idade ≥ 18 anos que realizaram colposcopia no departamento de Prevenção de Câncer no Hospital do Câncer de Barretos/SP no período de 2017 a 2019. RESULTADOS: Foram incluídas 3.411 mulheres, sendo 58,0% positivas para HPV de alto risco, e maior prevalência de NIC2+ para HPV16 (30,3%) e outros HPV (45,0%). Resultados citológicos sugestivos de lesões invasivas (epidermoide ou adenocarcinoma), independente do teste de HPV, apresentaram 100% de diagnóstico NIC2 + , enquanto atipias sugestivas de lesões de alto grau, HSIL e ASC-H, associados a HPV positivo, apresentaram 86 e 55,2%, respectivamente. Resultados citológicos de ASC-H entre mulheres > 40 anos e HPV negativo foram associados principalmente a achados benignos. Observamos que ≤ NIC1 apresenta uma maior prevalência entre mulheres mais velhas com HPV negativo, enquanto para lesões de alto grau, há um aumento entre mulheres mais jovens positivas para HPV16/18. Para diagnóstico de câncer, observamos que há um predomínio de HPV16/18 independente da faixa etária. CONCLUSãO: Foi identificado maior risco de lesões precursoras e câncer entre mulheres com HPV 16/18 positivo e atipias citológicas graves em testes de rastreio populacional. Além disso, resultados citológicos de ASC-H quando associados a HPV negativo com idade > 40 anos habitualmente representam achados benignos em investigação histológica.

Identification and performance evaluation of housekeeping genes for microRNA expression normalization by reverse transcription-quantitative PCR using liquid-based cervical cytology samples.

Screening for cervical cancer by cytology has been effective in reducing the worldwide incidence and mortality rates of this disease. However, a number of studies have demonstrated that the sensitivity of conventional cervical cytology may be too low for detection of cervical intraepithelial neoplasias (CIN). Therefore, it is important to incorporate more sensitive molecular diagnostic tests that could substantially improve the detection rates and accuracy for identifying CIN lesions. MicroRNAs (miRNAs) are a class of small non-coding RNAs with the potential to provide robust non-invasive cancer biomarkers for detecting CIN lesions in liquid-based cervical cytology (LBC) samples. At present, there is no consensus on which are the best housekeeping genes for miRNA normalization in LBC. The present study aimed to identify housekeeping genes with consistent and reproducible performance for normalization of reverse transcription-quantitative PCR (RT-qPCR) expression analysis of miRNA using LBC samples. The present study firstly selected six potential candidate housekeeping genes based on a systematic literature evaluation. Subsequently, the expression levels of microRNAs U6, RNU-44, RNU-47, RNU-48, RNU-49 and hsa-miR-16 were measured in 40 LBC samples using RT-qPCR. The stability of each potential housekeeping gene was assessed using the NormFinder algorithm. The results revealed that U6 and RNU-49 were the most stable genes among all candidates requiring fewer amplification cycles and smaller variation across the sample set. However, RNU-44, RNU-47, RNU-48 and hsa-miR-16 stability exceeded the recommended housekeeping value suitable for normalization. The findings revealed that U6 may be a reliable housekeeping gene for normalization of miRNA RT-qPCR expression analysis using LBC samples.

Risk Profile of High-grade Cervical Lesions and Cervical Cancer Considering the Combination of Cytology, HPV Genotype, and Age among Women Undergoing Colposcopy / Perfil de risco de lesões cervicais de alto grau e câncer cervical considerando a combinação de citologia, genótipo do HPV e idade entre mulheres submetidas a colposcopia

Abstract Objective The present study aims to establish a risk profile for high-grade cervical lesions and cervical cancer (CIN2 + ) in women undergoing colposcopy at the Hospital do Câncer de Barretos, through the analysis of Human Papillomavirus (HPV) infection, cervical cytology, and patient's age. Methods Retrospective cross-sectional study based on a computerized database of women aged ≥ 18 years old who underwent colposcopy at the Prevention Department of the Hospital do Câncer de Barretos from 2017 to 2019. Results A total of 3,411 women were included, 58.0% were positive for high-risk-HPV test, with a higher prevalence of CIN2+ for HPV16 (30.3%) and other HPV (45.0%). Cytological findings that suggest invasive cervical cancer (squamous cells or adenocarcinoma), regardless of the status of HPV test, showed 100% diagnosis of CIN2 + , while atypias that suggest high-grade lesions, HSIL and ASC-H, positive for HPV test, showed in 86 and 55.2%, respectively, diagnosis of CIN2 + . ASC-H cytological results among women aged > 40 years old and negative HPV were mainly associated with benign findings. We observed that ≤ CIN1 has a higher prevalence among older women with negative HPV, while for high-grade lesions there is an increase among young women HPV16- and/or 18-positive. In cancer diagnosis, we observed a predominance of HPV 16/18 regardless of the age group. Conclusion The highest risks of precursor lesions and cervical cancer were found among women with positive HPV 16/18 tests and severe cytological atypia in population screening tests. In addition, cytological findings of ASC-H HPV negative in women > 40 years old usually represent benign findings in histological investigation.

Resumo Objetivo Estabelecer um perfil de risco de lesões intraepiteliais de alto grau e câncer do colo do útero (NIC2 + ) em mulheres submetidas a colposcopia considerando-se a infecção pelo papilomavírus humano (HPV), citologia cervical e idade. Métodos Estudo retrospectivo transversal em banco de dados informatizado de mulheres com idade ≥ 18 anos que realizaram colposcopia no departamento de Prevenção de Câncer no Hospital do Câncer de Barretos/SP no período de 2017 a 2019. Resultados Foram incluídas 3.411 mulheres, sendo 58,0% positivas para HPV de alto risco, e maior prevalência de NIC2+ para HPV16 (30,3%) e outros HPV (45,0%). Resultados citológicos sugestivos de lesões invasivas (epidermoide ou adenocarcinoma), independente do teste de HPV, apresentaram 100% de diagnóstico NIC2 + , enquanto atipias sugestivas de lesões de alto grau, HSIL e ASC-H, associados a HPV positivo, apresentaram 86 e 55,2%, respectivamente. Resultados citológicos de ASC-H entre mulheres > 40 anos e HPV negativo foram associados principalmente a achados benignos. Observamos que ≤ NIC1 apresenta uma maior prevalência entre mulheres mais velhas com HPV negativo, enquanto para lesões de alto grau, há um aumento entre mulheres mais jovens positivas para HPV16/18. Para diagnóstico de câncer, observamos que há um predomínio de HPV16/18 independente da faixa etária. Conclusão Foi identificado maior risco de lesões precursoras e câncer entre mulheres com HPV 16/18 positivo e atipias citológicas graves em testes de rastreio populacional. Além disso, resultados citológicos de ASC-H quando associados a HPV negativo com idade > 40 anos habitualmente representam achados benignos em investigação histológica.