¿Está la virtud en el punto medio? La incertidumbre del CIN2 y el papel del VPH / Is virtue at the midpoint? The uncertainty of CIN2 and the role of HPV

Objetivo: La biopsia guiada por colposcopia (BGC) marca el manejo de la neoplasia intraepitelial cervical. El objetivo de este estudio fue evaluar la concordancia de los resultados entre la BGC y la escisión amplia de la zona de transformación (LLETZ, large loop excision of the transformation zone), y la utilidad del genotipado del virus del papiloma humano (VPH) para seleccionar a las pacientes con riesgo de lesión intraepitelial escamosa de alto grado/neoplasia intraepitelial cervical 3 (HSIL/CIN3). Método: Se compararon los resultados de la BGC y de la LLETZ, siendo esta última el método de referencia. Se evaluó la relación del genotipo del VPH con el diagnóstico final de HSIL/CIN3. Resultados: La precisión de la biopsia comparada con LLETZ fue del 61,4%. La tasa de concordancia fue del 64,4% para CIN1, del 31,4% para CIN2 y del 77,4% para CIN3. La tasa global de sobrediagnóstico fue del 18,68% y la de subdiagnóstico del 19,89%. En mujeres menores de 30 años, la concordancia fue del 62,79% (CIN1 65%, CIN2 39,58% y CIN3 73,08%), la tasa de sobrediagnóstico del 22,67% y la tasa de subdiagnóstico del 15,11%. La infección por VPH16 tuvo una odds ratio de 3,86 para el diagnóstico final de HSIL/CIN3+. Conclusiones: El diagnóstico de CIN2 por BGC parece insuficiente para seleccionar a las pacientes para tratamiento escisional, principalmente en mujeres jóvenes. El hallazgo de VPH16 es un factor de riesgo de HSIL/CIN3+ independientemente del resultado de la biopsia.

Objective: Colposcopy-guided biopsy (CGB) is a basic tool for the management of cervical intraepithelial neoplasia. The aim of this study is to evaluate the concordance of results between CGB and large loop excision of the transformation zone (LLETZ), and the usefulness of human papillomavirus (HPV) genotyping to select patients at risk of H-SIL/CIN3. Method: The results of colposcopy-guided biopsy and LLETZ were compared, with LLETZ being the gold standard. The relationship of HPV genotype to the final diagnosis of CIN3 was assessed. Results: The accuracy of CGB compared to LLETZ was 61.4%. The concordance rate was 64.4% for CIN1, 31.4% for CIN2 and 77.4% for CIN3. The overall overdiagnosis rate was 18.68% and underdiagnosis rate was 19.89%. In women under 30 years of age the concordance rate was 62.79% (CIN1 65%, CIN2 39.58% and CIN3 73.08%), and the rate of overdiagnosis and underdiagnosis was 22.67% and 15.11%, respectively. HPV16 infection had an odds ratio of 3.86 for the final diagnosis of CIN3+ and the result was significant regardless of the biopsy result. Conclusions: The CGB result as CIN2 is inaccurate and seems insufficient to select patients for excisional treatment, mainly in young women. HPV16 infection is a risk factor for CIN3+ regardless of the colposcopy-guided biopsy result.

Influence of IL-6, IL-8, and TGF-β 1 gene polymorphisms on the risk of human papillomavirus-infection in women from Pernambuco, Brazil

Human papillomavirus (HPV) infections are strongly associated with the development of cervical intraepithelial neoplasias and invasive cervical cancer. Polymorphisms in cytokine-encoding genes and behavioural cofactors could play an important role in protecting an individual against viral infections and cancer. Here, we investigated whether IL-6 -174 G>C, IL-8 +396 G>T, and TGF-β1 +869 G>C and +915 G>C polymorphisms were associated with susceptibility to HPV infection in women from north-east (Pernambuco) Brazil. We analysed 108 healthy uninfected women (HC) and 108 HPV-positive women with cervical lesions. Genetic polymorphisms were assessed using Sanger sequencing and polymerase chain reaction-restriction fragment length polymorphism. Comparison of the distribution of the genotypic and allelic frequencies of the IL-18 +396 T>G polymorphism between HPV infected woman an uninfected controls showed that the GG genotype and G allele were both more frequent in the HC group, and were associated with protection from HPV infection (p = 0.0015; OR = 0.29 CI95% = 0.13-0.61; p = 0.0005; OR = 0.45 CI95% 0.29-0.7, respectively). Individuals from the control group could have previously had HPV infection that was spontaneously eliminated; however, it was undetectable at the time of sample collection. Based on our findings, we hypothesize that the IL-8 +396 G>T polymorphism could interfere with susceptibility to HPV infection, by modulating the ability of immune system to fight the virus.

High Risk HPV E6/E7 Oncoprotein Expression in Women with High Grade Squamous Intraepithelial Lesion / Expressão de oncoproteinas E6/e7 do HPV de alto risco em Mulheres com Lesão Intraepitelial Escamosa de alto grau

Purpose To correlate the expression of high-risk HPV E6 mRNA with pap smear, colposcopy, and biopsy results in women with high grade squamous intraepithelial lesion (HSIL). Methods A cross-sectional study was performed on women referred for primary care services after cytological diagnosis of HSIL. We evaluated the expression of E6/E7 mRNA of HPV types 16,18,31,33, and 45 and correlated the results with those of Pap smear, colposcopy, and biopsy. For amplification/detection of mRNA E6 / E7 we used NucliSENSEasyQ kit to detect HPV mRNA by polymerase chain reaction with primers/ probes for HPV types 16, 18, 31, 33, and 45. Results Out of 128 valid tests, the results of 30 (23.4%) tests were negative and 98 (70%) tests were positive. Only one type of HPV was detected in 87.7% of the E6/E7 mRNA positive cases. HPV16 was detected in 61.2% of the cases, followed by HPV33 (26.5%), HPV31 (17.3%), HPV18 (10%), and HPV45 (4.08%). Pap smear tests revealed that the E6/E7 test was positive in 107 (83.8%) women with atypical squamous cells - high grade (ASC-H), HSIL, or higher. The E6/E7 test was positive in 69 (57.5%) specimens presenting negative cytology results. When analyzing the association with colposcopy results, the frequency of positive E6/E7 results increased with the severity of the injury, ranging from 57.1% in women without colposcopy-detected injury to 86.5% in those with higher levels of colposcopy findings. Of the 111 women who underwent biopsy and E6/E7 testing, the E6/E7 test was positive in 84.7% of the women who presented with lesions of cervical intraepithelial neoplasia (CIN) grade 2 or higher. Finally, 41.2% of women with a negative biopsy presented a positive E6/E7 test. Conclusions E6/E7mRNA expression was higher in women with HSIL and CIN grade 2 or higher.

Objetivo Correlacionar a expressão mRNAE6/E7 do HPV de alto risco com os exames de Papanicolau, colposcopia e biópsia em mulheres com lesão intraepitelial escamosa de alto grau (HSIL). Métodos Estudo transversal com mulheres encaminhadas aos serviços de atenção primária com diagnóstico citológico de HSIL. Foi avaliada a expressão do mRNAE6/E7 dos tipos de HPV 16,18,31,33 e 45, correlacionando-se a expressão com os exames de Papanicolau, colposcopia e biópsia. Para a amplificação/detecção demRNA de E6/E7 foi usado o kit NucliSENS EasyQ(r) HPV que detecta mRNA do HPV por meio da reação em cadeia da polimerase com primers/probes HPV dos tipos 16, 18, 31, 33 e 45. Resultados Foram obtidos 128 testes válidos. Destes: 30 (23,4%) foram negativos e 98 (70%) dos testes foram positivos. Foi encontrado apenas um tipo de HPV em 87,7% dos positivos. O HPV16 foi omais encontrado em61,2%, seguido pelos HPV33 (26,5%); HPV31 (17,34%); HPV 18 (10,0%) e HPV (45 4,0%). Quanto ao exame de Papanicolau, o teste E6/E7 foi positivo em 107 (83,8%) das mulheres com ASC-H, HSIL ou superior, enquanto em citologia negativa foi encontrado um resultado positivo em 69 (57,5%) colposcopia. A frequência de teste E6/E7 positivo aumentou com a gravidade da lesão, detectada na colposcopia variando de 57,1% em mulheres sem lesão identificada em colposcopia até 86,5% naqueles com achado de colposcopia de grau maior. Das 111 mulheres que se submeteram a biópsia e o teste E6/E7, o teste foi positivo em 84,7% das que apresentaramlesão igual ou superior a NIC 2 (neoplasia intraepitelial cervical) e 41,2% daqueles com biópsia negativa. Conclusões A expressão de E6, E7 RNAm ocorreu commaior frequência emlesões de alto grau citológica e em casos com biópsias de NIC2 ou maior.

CCR2 and CCR5 genes polymorphisms in women with cervical lesions from Pernambuco, Northeast Region of Brazil: a case-control study

Polymorphisms in chemokine receptors play an important role in the progression of cervical intraepithelial neoplasia (CIN) to cervical cancer (CC). Our study examined the association of CCR2-64I (rs1799864) andCCR5-Δ32 (rs333) polymorphisms with susceptibility to develop cervical lesion (CIN and CC) in a Brazilian population. The genotyping of 139 women with cervical lesions and 151 women without cervical lesions for the CCR2-64I and CCR5-Δ32 polymorphisms were performed using polymerase chain reaction-restriction fragment length polymorphism. The individuals carrying heterozygous or homozygous genotypes (GA+AA) for CCR2-64I polymorphisms seem to be at lower risk for cervical lesion [odds ratio (OR) = 0.37, p = 0.0008)]. The same was observed for the A allele (OR = 0.39, p = 0.0002), while no association was detected (p > 0.05) with CCR5-Δ32 polymorphism. Regarding the human papillomavirus (HPV) type, patients carrying the CCR2-64Ipolymorphism were protected against infection by HPV type 16 (OR = 0.35, p = 0.0184). In summary, our study showed a protective effect ofCCR2-64I rs1799864 polymorphism against the development of cervical lesions (CIN and CC) and in the susceptibility of HPV 16 infection.

An upward trendiin dna p16ink4a methylation patern and high risk hpv infectionn according to the severity of the cervical lesion / Padrão de metilação do gene P16INK4A e de infecções por HPV de alto risco acompanham a evolução de lesões cervicais

SUMMARY High-risk human papillomavirus (hr-HPV) infection is necessary but not sufficient for cervical cancer development. Recently, P16INK4A gene silencing through hypermethylation has been proposed as an important cofactor in cervical carcinogenesis due to its tumor suppressor function. We aimed to investigate P16INK4A methylation status in normal and neoplastic epithelia and evaluate an association with HPV infection and genotype. This cross-sectional study was performed with 141 cervical samples from patients attending Hospital Moncorvo Filho, Rio de Janeiro. HPV detection and genotyping were performed through PCR and P16INK4A methylation by nested-methylation specific PCR (MSP). HPV frequency was 62.4% (88/141). The most common HPV were HPV16 (37%), HPV18 (16.3%) and HPV33/45(15.2%). An upward trend was observed concerning P16INK4A methylation and lesion degree: normal epithelia (10.7%), low grade lesions (22.9%), high grade (57.1%) and carcinoma (93.1%) (p < 0.0001). A multivariate analysis was performed to evaluate an association between methylation, age, tobacco exposure, HPV infection and genotyping. A correlation was found concerning methylation with HPV infection (p < 0.0001), hr-HPV (p = 0.01), HSIL (p < 0.0007) and malignant lesions (p < 0.0001). Since viral infection and epigenetic alterations are related to cervical carcinoma, we suggest that P16INK4A methylation profile maybe thoroughly investigated as a biomarker to identify patients at risk of cancer. .

RESUMO É reconhecido que infecções por papilomavírus humanos de alto risco (HPV) são causa necessária, mas não suficiente para o desenvolvimento do câncer cervical. Recentemente, estudos de silenciamento gênico apontaram que a hipermetilação do gene p16INK4A é importante co-fator para a carcinogênese cervical, eliminando a função supressora de tumor da proteína p16 em lesões malignas. Entretanto poucos estudos avaliaram a relação da metilação com a progressão da doença. Nosso objetivo foi investigar o padrão de metilação do gene P16INK4A em diferentes graus de lesão cervical e sua associação com a infecção por diferentes tipos de HPV. Nosso estudo de corte transversal avaliou 141 amostras cervicais de pacientes atendidas no Hospital Moncorvo Filho, Rio de Janeiro. A detecção e tipagem do HPV foi realizada pela técnica de reação em cadeia da polimerase (PCR), e a metilação do gene P16INK4A pela PCR-metilação específica em formato nested (MSP). A frequência de HPV foi de 62,4% (88/141). O tipo mais prevalente foi o HPV16 (37%), seguido pelo HPV18 (16,3%) e HPV33/45 (15,2%). Curva ascendente foi observada quanto ao padrão de metilação do gene P16INK4A e o grau da lesão: a metilação foi identificada em somente 10,7% das amostras de epitélio normal, em 22,9% das lesões de baixo grau, em 57,1% das lesões de alto grau e em 93,1% dos carcinomas (p < 0,0001). Foram feitas análises univariada e multivariada a fim de correlacionar metilação, idade, exposição ao tabaco, infecção e genótipo de HPV. Foi encontrada correlação da metilação com a infecção pelo HPV (p < 0,0001), genótipos de alto risco (p = 0,01), ...

Metilação genética, neoplasia intraepitelial cervical e câncer do colo uterino / Genetics methylation, cervical intraepithelial neoplasia and cervical cancer

O câncer de colo uterino é uma importante causa de morte entre as mulheres em países subdesenvolvidos. A infecção persistente pelo papilomavírus humano (HPV) oncogênico e o comprometimento da resposta imune são fatores de risco para o desenvolvimento da neoplasia intraepitelial cervical (NIC) e sua progressão para o câncer cervical invasivo. O diagnóstico precoce e o tratamento das lesões precursoras do câncer são de grande importância. Estudos epigenéticos estão sendo realizados com o objetivo de avaliar sua influencia nos processos de oncogênese, visto que alterações epigenéticas estão presentes em quase todos os tumores. A metilação de DNA e a acetilação de histonas são as duas mudanças epigenéticas mais estudadas. O melhor entendimento do perfil epigenético na neoplasia intraepitelial cervical e no câncer cervical invasor pode ser utilizado no diagnóstico e prognóstico deste câncer. O objetivo desta revisão consistiu em entender as mudanças epigenéticas encontradas até o momento nas pacientes com NIC e câncer de colo uterino. Foi realizada revisão da literatura de estudos indexados em banco de dados, como PubMed e LILACS. Verificou-se que, até o presente momento, não há um marcador de metilação que tenha o desempenho adequado para servir como indicador para as lesões precursoras do câncer, ou mesmo para o carcinoma cervical.

The cervical cancer is a major cause of death among women in developing countries. Persistent infection by human papillomavirus (HPV) and oncogenic involvement of the immune response are risk factors for the development of cervical intraepithelial neoplasia (CIN) and its progression to invasive cervical cancer. Early diagnosis and treatment of cancer precursor lesions are of great importance. Epigenetic studies are being conducted to evaluate its influence in the process of oncogenesis, since epigenetic alterations are present in almost all tumors. DNA methylation and histone acetylation are the two most studied epigenetic changes. An improved understanding of the epigenetic profile in CIN and invasive cervical cancer can be used in the diagnosis and prognosis of this cancer. The aim of this review was to understand the epigenetic changes found to date in patients with CIN and cervical cancer. We performed a literature review of studies indexed in databases such as PubMed and LILACS. It was found that, to our knowledge, there is no methylation marker with an adequate performance to serve as an indicator for cancer precursor lesions, or even for cervical carcinoma.

Mecanismo de infección y transformación neoplásica producido por virus papiloma humano en el epitelio cervical / Infection mechanism and neoplastic transformation produced by human papilloma virus in cervical epithelium

Se presenta una revisión bibliográfica del mecanismo de infección y transformación neoplásica producida por el virus papiloma humano, de alto riesgo oncogénico, en el epitelio cervical. Se expone la interacción cápside receptor, internalización celular, expresión de genes tempranos, integración del genoma viral al de la célula huésped y algunos mecanismos vinculados a la proliferación y desarrollo neoplásico.

Chromosome 3p alterations in northeastern Thai women with cervical carcinoma.

The purpose of this study was to determine the incidence of the loss of heterozygosity (LOH) among normal cervixes, cervical intraepithelial neoplasias (CINs) and invasive cervical cancers (ICCs). DNA samples (136) were obtained from 31 normal cervixes, 49 CINs and 56 ICCs. Four polymorphic microsatellite markers (D3S1300, D3S1351, D3S1478 and D3S4103) covering the chromosome 3p arm, were employed. LOH at one or more loci were identified in: 9/31 (8.1%) normal cervixes, 17/49 (14.6%) CINs and 26/56 (22.1%) invasive cancers. The incidence of the LOH at 3p varied for each locus and ranged from 5.6% for D3S1351 to the highest rate of 16.6% for D3S1300. We thus found that LOH of chromosome 3p can occur in normal cervixes and that incidences increase in CINs and ICCs. Deletion in the 3p14.2 (D3S1300) and 3p21.2 (D3S1478) regions might be an early event and, in fact, necessary for cervical cancer progression. The loss of function of tumor suppressor genes (TSGs) located in these regions may have a sequential effect in cervical cancer carcinogenesis.

Predictive value of morphological nuclear parameters and DNA ploidy pattern in precancerous lesions of the uterine cervix.

To evaluate the supportive role of image cytometry and DNA ploidy analysis in the precancerous and cancerous lesions of the uterine cervix, the present study was performed on 45 cervical smears, initially diagnosed as dysplasia and malignant. Twenty normal and inflammatory smears were taken as a control for the study. Morphometric parameters and microphotometric DNA measurements were performed on 50 cells in each case. On the basis of nuclear area dysplastic lesions were categorised into two groups i.e. low grade lesions having nuclear area upto 85 sq. mu m and high grade lesions having nuclear area above 85 sq. mu m. The results were compared with DNA ploidy analysis. It is revealed from the study that 85.7% low grade lesions with diploid and polyploid DNA, value mostly regressed to inflammation and 78.5% high grade lesion with aneuploid DNA value progressed to malignancy. However, initial malignant cases having aneuploid DNA value exhibit invasive cancer during their follow up. It indicates that combination of morphometry and DNA cytometry can be used as an adjunct to cytologic diagnosis to predict the biologic outcome of the lesions.

Detección del papilomavirus humano en ADN de tejido normal y lesiones premalignas del cérvix por hibridación molecular / Identification of human papilloma virus in DNA of normal tissue and precursor lesions of uterine cervix by molecular hybridization

Se analizaron 64 muestras de ADN obtenido de tejido histológicamente normal del cérvix y de lesiones tempranas del cáncer cervical por hibridación dot-blot, para detectar la presencia de papilomavirus humano(PVH). Utilizando ADN de controles adecuados y sondas específicas, detectamos la infección por PVH en 39 por ciento(25 de 64) de los casos; mientras que 61 por ciento (39 de 64) resultó negativo a PVH. Se detectó PVH del grupo de ®bajo riesgo¼ (tipo 6/11) en un 80 por ciento (20 de 25) de las muestras de ADN; mientras que un 8 por ciento (2 de 25) fue positivo a PVH del grupo de ®alto riesgo¼ (tipo 16/18). Además, encontramos 12 por ciento (3 de 25) de las muestras positivas a ambos grupos de virus

Correlación citológica e histológica de las imágenes colposcópicas en el diagnóstico de neoplasia cervical

Se revisaron las historias clínicas de las pacientes de la clínica de colposcopia, que ingresaron en el estudio entre los años de 1988-1989. Se estudiaron las imágenes colposcópicas y se correlacionaron con los diferentes estudios: Citología, resultado de patología en la biopsia y en la pieza final. El estudio incluye 400 casos. La sensibilidad de la citología corresponde al 93 por ciento, especificidad el 42 por ciento y falsos negativos del 7 por ciento. La citología fue negativa en 4 por ciento de los casos con NIC III y en 9 por ciento de los casos con Ca. invasivo. La citología A2 con atipias severas se correlacionó con casos de NIC II -III en 21 por ciento de las pacientes. La citología A5 presenta una frecuencia de falsos negativos par Ca Invasivo de 44 por ciento y la citología A4-A5, presenta tasas de falsos negativos frente a cérvix del 76 por ciento. El porcentaje de colposcopia insatisfactoria es del 34 por ciento, siendo en mayores de 40 años de 52 por ciento. Las pacientes con lesión endocervical presentan una tasa de colposcopia insatisfactoria del 67 por ciento. La sensibilidad de la colposcopia en nuestra institución es del 88 por ciento con una especificidad del 39 por ciento y unos falsos negativos del 12 por ciento, frente a la citología. Frente al resultado de patología, la colposcopia tiene una sensibilidad del 89 por ciento, una especificidad del 64 por ciento y una tasa de falsos negativos del 11 por ciento. En el estudio de lesión por papilomavirus, la citología solo da una sensibilidad del 12 por ciento, con un índice de falsos negativos del 88 por ciento. La colposcopia frente a la lesión por condiloma da una sensibilidad del 63 por ciento y unos falsos negativos del 37 por ciento. En los pacientes con biopsia que refería compromiso glandular, la anatomía patológica final no reveló en ningún caso invasión. La correlación de imágenes colposcópicas se establece así: Cervicitis: zona de transformación típica, epitelio acetoblanco

Human papillomavirus detection in cervical dysplasias or neoplasias and in condylomata acuminata by in situ hybridization with biotinylated DNA probes

Specimens from cervical dysplasias or carcinomas and genital condylomata acuminata were retrospectively analysed by in situ hybridization (ISH) with biotinylated DNA probes for human papillomavirus (HPV) types 6, 11, 16 and 18. In the control group no case was positive for HPV DNA. In mild/moderate dysplasias, 4 cases (14%) were positive for HPV 6 or 11 and 2 cases (7%), for HPV 16. In the severe dysplasia/in situ carcinoma group, 9 cases (31%) showed presence of DNA of HPV types 16 or 18. Six invasive carcinomas (20%) were positive for HPV type 16 or 18. Among condylomata acuminata, 22 cases (73%) were positive for HPV types 6 or 11. In all ISH-positive cases only one viral type was detected. No correlation between HPV DNA positivity and histological findings of HPV infection was observed. Although less sensitive than some other molecular biology techniques, in situ hybridization with biotinylated DNA probes proved to be simple and useful for detecting and typing HPV in samples routinely received for histopathological analysis