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Brachybacterium conglomeratum, traditionally considered an environmental bacterium, has recently garnered attention for its potential involvement in human health. While prior research hinted at its pathogenic role in humans, our study aims to determine its prevalence and associations in diverse clinical contexts. We examined vaginal swabs from three distinct patient groups: patients with low-grade squamous intraepithelial lesions (LSIL), patients with cervicovaginal infections, and patients with a history of precancerous lesions undergoing follow-up. B. conglomeratum was present in all three patient groups, with the highest prevalence observed in the LSIL group. Statistically significant associations were primarily identified in the LSIL group, where B. conglomeratum was present in 60% of cases. Notably, the LSIL group exhibited coinfections with multiple high-risk oncogenotypes of human papillomavirus (HPV), suggesting potential synergistic effects, and understanding these microbial relationships and their influence on viral persistence, particularly with HPV, holds promise for mitigating HPV-related carcinogenesis. Furthermore, Gardnerella vaginalis and Atopobium vaginae were frequently detected in this group, along with Ureaplasma parvum as the predominant sexually transmitted bacterium. In all cases, B. conglomeratum was found in association with these microorganisms rather than as a sole pathogen. This coexistence underscores the intricate microbial interactions within cervicovaginal infections and precancerous lesions. This study marks the first report of B. conglomeratum prevalence in women with these clinical conditions.
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The relationship among microbiome, immunity and cervical cancer has been targeted by several studies, yet many questions remain unanswered. We characterized herein the virome and bacteriome from cervical samples and correlated these findings with innate immunity gene expression in a Brazilian convenience sample of HPV-infected (HPV+) and uninfected (HPV-) women. For this purpose, innate immune gene expression data were correlated to metagenomic information. Correlation analysis showed that interferon (IFN) is able to differentially modulate pattern recognition receptors (PRRs) expression based on HPV status. Virome analysis indicated that HPV infection correlates to the presence of Anellovirus (AV) and seven complete HPV genomes were assembled. Bacteriome results unveiled that vaginal community state types (CST) distribution was independent of HPV or AV status, although bacterial phyla distribution differed between groups. Furthermore, TLR3 and IFNαR2 levels were higher in the Lactobacillus no iners-dominated mucosa and we detected correlations among RIG-like receptors (RLR) associated genes and abundance of specific anaerobic bacteria. Collectively, our data show an intriguing connection between HPV and AV infections that could foster cervical cancer development. Besides that, TLR3 and IFNαR2 seem to create a protective milieu in healthy cervical mucosa (L. no iners-dominated), and RLRs, known to recognize viral RNA, were correlated to anaerobic bacteria suggesting that they might be related to dysbiosis.
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Microbiota , Infecciones por Papillomavirus , Neoplasias del Cuello Uterino , Femenino , Humanos , Cuello del Útero , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/metabolismo , Brasil , Receptor Toll-Like 3/genética , Bacterias/genética , Expresión GénicaRESUMEN
Introducción: El cáncer cervicouterino inicia con una lesión precancerosa llamada displasia, pudiendo ser de bajo grado o alto grado; uno de los factores más importantes en este sentido es la edad de inicio de la vida sexual activa (IVSA). Objetivo: Conocer si existe asociación entre la edad de inicio de la vida sexual activa y la lesión intraepitelial escamosa de alto grado (LIEAG). Material y métodos: Estudio descriptivo, retrospectivo, observacional, en 52 expedientes de mujeres de 15 a 60 años, con Papanicolaou en el HGZ MF No. 1, previa autorización del CLIS 301 y el CEI 3018, con folio R-2022-301-021. Se recolectaron los datos de expedientes que cumplieron los criterios de inclusión; se realizó un análisis estadístico con frecuencias, porcentajes para variables cualitativas nominales, con medidas de tendencia central y dispersión en variables cuantitativas, con determinación de chi2 y prueba de Kruskal-Wallis, respetándose los principios de Belmont en consideración a la Justicia y Beneficencia, los principios éticos de la Declaración de Helsinki 1964 y la Ley General de Salud en México. Resultado: Al evaluar 52 expedientes de pacientes, la edad fue de 37.81 ± 9.949 años; se encontró asociación entre el IVSA y la LIEAG, sin significación estadística, de acuerdo con la prueba de chi2, con un valor de p = 0.538 (IC 95%: 0.403-0.674), y una asociación significativa por la prueba de Kruskal-Wallis entre la LIEAG y la edad de la paciente, con un valor de p = 0.019 (IC 95%: 0.000-0.057). Conclusión: El IVSA no se correlaciona con el tipo de LIEAG.
Introduction: Cervical cancer begins with a precancerous lesion called dysplasia, which can be low grade or high grade. One of the most important factors is the age at which an active sexual life begins. Objective: To know if there is an association between the age of beginning of active sexual life (BASL) and high-grade squamous intraepithelial lesions (HGSIEL). Material and methods: Descriptive, retrospective, observational study, in 52 files of women aged 15 to 60 years with Pap smear at HGZ MF No. 1, prior authorization from CLIS 301 and CEI 3018 with folio R-2022-301-021. Data were collected from files that met the inclusion criteria, a statistical analysis was carried out with frequencies, percentages for nominal qualitative variables, with measures of central tendency and dispersion in quantitative variables, with determination of chi2 and Kruskal-Wallis, respecting the principles of Belmont in consideration of Justice and Beneficence, the ethical principles of the Declaration of Helsinki 1964, the General Health Law in México. Result: When evaluating 52 patient records, the age was 37.81 ± 9.949 years, the association of BASL and HGSIEL was found without statistical significance according to the chi2 with a p = 0.538 (95% CI, 0.403-0.674) and a significant association by Kruskal-Wallis between HGSIEL and patient age, with p = 0.019 (95% CI, 0.000-0.057). Conclusion: The beginning of an active sexual life has no association with the type of high-grade squamous intraepithelial lesions.
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Neoplasias del Cuello Uterino , Conducta Sexual , Estadísticas no Paramétricas , Lesiones Intraepiteliales EscamosasRESUMEN
Abstract Objective To determine the prevalence and possible variables associated with anal intraepithelial neoplasia and anal cancer in immunocompetent women with high-grade cervical intraepithelial neoplasia. Methods A cross-sectional study involving immunocompetent women with a histological diagnosis of high-grade cervical intraepithelial neoplasia and cervical cancer, conducted between January 2016 and September 2020. All women underwent anal cytology and answered a questionnaire on characterization and potential risk factors. Women with altered cytology were submitted to anoscopy and biopsy. Results A total of 69 women were included in the study. Of these, 7 (10.1%) had abnormal anal cytology results: (high-grade lesion, atypical squamous cells of undetermined significance, and atypical squamous cells, cannot exclude high-grade lesions: 28,5% each; low grade lesion: 14,3%). Of the anoscopies, 3 (42.8%) showed alterations. Of the 2 (28,5% of all abnormal cytology results) biopsies performed, only 1 showed low-grade anal intraepithelial neoplasia. The average number of pregnancies, vaginal deliveries, and abortions was associated with abnormal anal cytology. However, the highest mean regarding the cesarean sections was associated with normal cytology. Conclusion The prevalence of anal intraepithelial neoplasia was compatible with data from recent studies, especially those conducted in Brazil. Opportunistic screening for anal intraepithelial neoplasia in this high-risk population should be considered. Anal cytology is suitable for this purpose, due to its low cost and feasibility in public health services.
Resumo Objetivo Determinar a prevalência e as possíveis variáveis associadas à neoplasia intraepitelial anal e ao câncer anal em mulheres imunocompetentes com neoplasia intraepitelial cervical de alto grau. Métodos Estudo transversal em mulheres imunocompetentes com diagnóstico histológico de neoplasia intraepitelial cervical de alto grau e câncer cervical, feito entre janeiro de 2016 e setembro de 2020. Todas as mulheres foram submetidas a citologia anal e responderam a um questionário de caracterização e potenciais fatores de risco. Mulheres com citologia alterada foram submetidas a anuscopia e biópsia. Resultados No total, 69 mulheres foram incluídas no estudo. Destas, 7 (10,1%) tiveram resultados anormais de citologia anal (lesão de alto grau, células escamosas atípicas de significado indeterminado, e células escamosas atípicas, não se pode excluir lesões de alto grau: 28,5% cada; lesão de baixo grau: 14,3%). Das anuscopias, 3 (42,8%) demonstraram alterações. Das 2 biópsias realizadas, apenas 1 apresentou neoplasia intraepitelial anal de baixo grau. O número médio de gestações, partos vaginais e abortos estava associado à citologia anal anormal. No entanto, a maior média de partos cesáreos estava associada à citologia normal. Conclusão A prevalência de neoplasia intraepitelial anal foi compatível com dados de estudos recentes, principalmente daqueles feitos no Brasil. O rastreamento oportunista para neoplasia intraepitelial anal nesta população de alto risco deve ser considerado. A citologia anal é adequada para esse fim, devido ao seu baixo custo e viabilidade nos serviços públicos de saúde.
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Humanos , Femenino , Neoplasias del Ano/diagnóstico por imagen , Tamizaje Masivo , Lesiones Intraepiteliales Escamosas , PapillomaviridaeRESUMEN
Objetivo: Comparar la presencia del Virus de Papiloma Humano (VPH) y de lesión Intraepitelial Cervical (LIE) en adolescentes embarazadas y no grávidas atendidas en la Maternidad Dr. Armando Castillo Plaza de Maracaibo, Venezuela. Método: Investigación comparativa con diseño no experimental transeccional y de campo; donde se incluyeron 46 adolescentes embarazadas (casos) y 46adolescentes no embarazadas (controles), escogidas mediante muestreo probabilístico aleatorio, a quienes se les realizó identificación de factores asociados a la patología, evaluación por citología cervicovaginal y Genotipificación del VPH por reacción en cadena de la polimerasa (PCR). Resultados: se encontró que 32,6% de las embarazadas presentaron LIE de bajo grado (VPH o NIC 1) respecto a 21,7% en las no grávidas, común riesgo dos veces mayor (OR [IC95%]= 2,44 [1,05-5,65]). El diagnóstico molecular resultó positivo en la mitad del total de la muestra, siendo mayor en las embarazadas (52,1 vs. 47,9p<0,05);predominado las infecciones pro genotipos de alto riesgo 47,8vs 30,5; p <0,05). El VPH 16 resulto el más prevalente entre las embarazadas (21,7%) y la co-infección por genotipos debajo riesgo (6-11) en las no grávidas (17,4%) Conclusiones: las embarazadas adolescentes presentan una mayor prevalencia de LIE e infección genital por VPH, asociado a un riesgo significativo del doble de probabilidad de presentar una LIE respecto a las adolescentes no grávidas(AU)
Aim: To compare the presence of Human PapillomaVirus (HPV) and Squamous Intraepithelial Lesion (SIL)in pregnant and non-pregnant adolescents treated at the "Maternidad Dr. Armando Castillo Plaza" in Maracaibo, Venezuela. Patients and Methods: A comparative research with non-experimental transectional and field design was performed; where 46 pregnant adolescents (cases) and 46 non-pregnant adolescents (controls) was included, chosen by random probability sampling, who under went identification off actors associated with the pathology, evaluation by pap-smearand HPV genotyping by chain reaction of polymerase (PCR). Results: It was found that 32.6% of pregnant women had lowgrade SIL ( HPV or CIN 1) compared to 21.7% in non-pregnant women, with a risk twice higher (OR [95% CI] = 2.44 [1.05-5.65]). thee molecular diagnosis was positive in half of the total sample, being higher in pregnant women (52.1 vs. 47.9p <0.05);infections with high-risk genotypes predominated 47.8 vs 30.5;p <0.05). HPV 16 was the most prevalent among pregnant women (21.7%) and co-infection by low-risk genotypes (6-11) in non-pregnant women (17.4%). Conclusions: adolescent pregnant women have a higher prevalence of LIE and genital HPV infection, associated with a significant risk of twice the probability of presenting an LIE compared to non-pregnant adolescents(AU)
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Humanos , Masculino , Adolescente , Embarazo en Adolescencia , Neoplasias del Cuello Uterino , Infecciones por Papillomavirus , Papillomaviridae , Enfermedades de Transmisión Sexual , Células Epiteliales , Biología CelularRESUMEN
Objectives: To compare morphological abnormalities on anal colposcopy against histology to determine anal high-grade squamous intraepithelial lesions (HSILs). Methods: This is a retrospective data assessment of HIV-negative and HIV-positive patients undergoing outpatient follow-up. The sample comprised 54 patients presenting acetowhite lesions on anal colposcopy. Acetowhite lesions were classified according to their morphology into punctation, verrucous, mosaic, ulcerated, or hypervascularized, and biopsies of these specimens were classified as anal HSIL, low-grade squamous intraepithelial lesion (LSIL), or normal. The data were analyzed using SPSS forWindows version 13.0 (SPSS Inc., Chicago, IL, USA). The results were analyzed using the nonparametric Mann-Whitney test, the Fisher exact test and the chi-squared parametric test. A 95% confidence interval (CI) was used and a level of significance <5% was adopted for all statistical tests. Results Fifty-four patients (50 males, 80% HIV+) with biopsied acetowhite lesions were assessed. There were 31 punctation lesions, 1 classified as HSIL (3.2%; 95%CI: 0- 40.0), 17 verrucous lesions, 3 HSIL (17.7%; 95%CI: 0-10.7), and 1 ulcerated, classified as HSIL (100%), and 4 mosaic and 1 atypical vessel lesion, all classified as LSIL. The results showed no association of presence of anal HSIL with positivity for HIV infection or with counts above or below 500/µl in HIV+patients. Statistical analysis was performed using the Mann-Whitney nonparametric test, the Fisher exact test, and the chi-squared parametric test. Conclusion: The comparison of morphological findings on anal colposcopy against histology revealed no morphological pattern suggesting anal HSIL. (AU)
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Humanos , Masculino , Femenino , Adolescente , Adulto , Persona de Mediana Edad , Canal Anal , Carcinoma in Situ/ultraestructura , Condiloma Acuminado , Papillomaviridae , Colonoscopía , VIH , Recuento de Linfocito CD4RESUMEN
BACKGROUND: High-risk human papillomavirus (HR-HPV) infection is the main cause of cervical cancer, but additional alterations are necessary for its development. Abnormal DNA methylation has an important role in the origin and dissemination of cervical cancer and other human tumors. In this work, we analyzed the methylation of eight genes (AJAP1, CDH1, CDH13, MAGI2, MGMT, MYOD1, RASSF1A and SOX17) that participate in several biological processes for the maintenance of cell normality. We analyzed DNA methylation by methylation-specific PCR (MSP) and HPV infection using the INNOLiPA genotyping kit in 59 samples diagnostic of normal cervical tissue (non-SIL), 107 low-grade squamous intraepithelial lesions (LSILs), 29 high-grade squamous intraepithelial lesions (HSILs) and 51 cervical cancers (CCs). RESULTS: We found that all samples of LSIL, HSIL, and CC were HPV-positive, and the genotypes with higher frequencies were 16, 18, 51 and 56. In general, the genes analyzed displayed a significant tendency toward an increase in methylation levels according to increasing cervical lesion severity, except for the CDH13 gene. High CpG island methylator phenotype (CIMP) was associated with a 50.6-fold (95% CI 4.72-2267.3)-increased risk of HSIL and a 122-fold risk of CC (95% CI 10.04-5349.7). CONCLUSIONS: We found that CIMP high was significantly associated with HSIL and CC risk. These results could indicate that CIMP together with HR-HPV infection and other factors participates in the development of HSIL and CC.
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Islas de CpG/genética , Metilación de ADN/genética , Predisposición Genética a la Enfermedad , Fenotipo , Lesiones Intraepiteliales Escamosas de Cuello Uterino/genética , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/fisiopatología , Adulto , Línea Celular , Femenino , Regulación Neoplásica de la Expresión Génica , Genotipo , Humanos , Queratinocitos , México , Persona de Mediana Edad , Factores de RiesgoRESUMEN
As neoplasias intraepiteliais cervicais correspondem a alterações identificadas por rastreamento citológico cervical e estudo histológico, pós-biópsia incisional guiada por colposcopia ou procedimento diagnóstico excisional. Podem ser tratadas com abordagens conservadoras e procedimentos excisionais. A vacinação anti-HPV e o tratamento excisional oportuno constituem, respectivamente, prevenção primária e secundária contra o câncer do colo uterino.(AU)
Cervical intraephitelial neoplasms correspond to changes identified by cervical citological screening and histological study, post-incisional biopsy guided by colposcopy or excisional diagnostic procedure. They can be treated with conservative approaches and excision procedures. Anti-HPV vaccination and timely excional treatment are primary and secondary prevention against cervical cancer, respectively.(AU)
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Humanos , Femenino , Cuello del Útero/citología , /cirugía , /diagnóstico , Lesiones Intraepiteliales Escamosas/cirugía , Lesiones Intraepiteliales Escamosas/diagnóstico , Lesiones Intraepiteliales Escamosas/diagnóstico por imagen , /diagnóstico por imagen , Colposcopía , Conización/instrumentación , Infecciones por Papillomavirus/patología , Ultrasonido Enfocado de Alta Intensidad de Ablación , HisterectomíaRESUMEN
Persistent infection with the human papillomavirus 16 (HPV 16) is the cause of half of all cervical carcinomas (CC) cases. Moreover, mutations in the oncoproteins E6 and E7 are associated with CC development. In this study, E6/E7 variants circulating in southern Mexico and their association with CC and its precursor lesions were evaluated. In total, 190 DNA samples were obtained from scrapes and cervical biopsies of women with HPV 16 out of which 61 are from patients with CC, 6 from patients with high-grade squamous intraepithelial lesions (HSIL), 68 from patients with low-grade squamous intraepithelial lesions (LSIL), and 55 from patients without intraepithelial lesions. For all E7 variants found, the E7-C732/C789/G795 variant (with three silent mutations) was associated with the highest risk of CC (odd ratio (OR) = 3.79, 95% confidence interval (CI) = 1.46-9.85). The analysis of E6/E7 bicistron conferred to AA-a*E7-C732/C789/G795 variants revealed the greatest increased risk of CC (OR = 110, 95% CI = 6.04-2001.3), followed by AA-c*E7-C732/C789/G795 and A176/G350*E7-p. These results highlight the importance of analyzing the combinations of E6/E7 variants in HPV 16 infection and suggest that AA-a*E7-C732/C789/G795, AA-c*E7-C732/C789/G795, and A176/G350*E7-p can be useful markers for predicting CC development.
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OBJECTIVE: Our main objective was to assess the association between the markers p16 and Ki-67 and recurrence of disease in patients previously treated for cervical high-grade squamous intraepithelial lesion (HSIL). DESIGN: This is a case-control study at the National Cancer Institute conducted between 2005 and 2015. Of the patients with a pathologically confirmed diagnosis of HSIL, 107 cases were selected. They were divided into 2 groups: 28 cases with recurrence after treatment and a control group of 79 patients without recurrence. We identified clinical, pathological, and treatment variables. METHODS: Two experienced pathologists performed immunohistochemical analysis of biomarkers; they agreed on their interpretation, and we calculated the odds ratios (ORs) associated with recurrence. For group comparisons, we used the Wilcoxon signed-rank, χ2, or Fisher's exact test, depending on the type of variable. We conducted logistic regression models to estimate ORs and determine the factors associated with recurrence. The recurrence-free period was defined as the time frame between conization and either recurrence of disease or the last date the patient was seen. We used Kaplan-Meier plots to visualize survival curves and log-rank tests to compare the curves. We established a p value <0.05 as statistically significant. RESULTS: After pathologists performed immunohistochemical analysis, they achieved an agreement level of 83.7% for p16 and 60% for Ki-67. We did not find an association between recurrence and either p16 expression (p = 0.69) or the percentage of Ki-67 expression (p = 0.71). The recurrence-free period analysis did not reveal a difference in p16 expression (p = 0.57) nor in the percentage of Ki-67 expression in the 3-tiered scale (p = 0.56). LIMITATIONS: Our main limitation was a reduced sample size. CONCLUSION: We found no association between p16 and Ki-67 positivity and the risk of recurrence in previously treated HSIL.
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Infecciones por Papillomavirus , Lesiones Intraepiteliales Escamosas de Cuello Uterino , Lesiones Intraepiteliales Escamosas , Displasia del Cuello del Útero , Neoplasias del Cuello Uterino , Biomarcadores de Tumor , Estudios de Casos y Controles , Inhibidor p16 de la Quinasa Dependiente de Ciclina , Femenino , Humanos , Antígeno Ki-67 , Recurrencia Local de NeoplasiaRESUMEN
BACKGROUND: Radiofrequency ablation (RFA) of high-grade squamous intraepithelial lesions (HSIL) is a promising minimally invasive technique but its oncologic and functional outcomes are not well studied. The primary outcome was the efficacy of RFA, and the secondary outcomes were the functional and anatomical anal changes related to RFA. METHODS: This was a retrospective analysis of our prospectively collected database of patients who had RFA for HSIL at our institution, between August 2018 and March 2020. To be eligible for RFA, all patients had impairment of their immune function. Targeted ablation was applied in all cases, with 5 overlapping pulsations at the targeted HSILs (delivering 12 J/cm2 per application) followed by circumferential, 2-pulsation (12 J/cm2) overlapping anal ablation, to cover the entire anal transition zone. Patients were assessed for recurrence or metachronous disease at 3-month intervals by means of high-resolution anoscopy (HRA) and targeted biopsies. Anorectal manometry, endoanal ultrasound, the 36-Item Short Form and Massachusetts General Hospital-Sexual Functioning Questionnaire (MGH-SFQ) were assessed at baseline and 12 months after intervention. RESULTS: We included a total of 12 patients with anal HSILs. The mean age was 38.6 (± 7.68) years, and 7 (58.3%) were males. Six were HIV positive, 2 had a primary immunodeficiency disease, and 4 were receiving immunosuppressive therapy. A mean of 2.1 anal HSILs per patient were treated. At 12 months, high-resolution anoscopy showed that 7/12 (58.3%) patients had normal high-resolution anoscopy, 3/12 patients had recurrent HSILs, and 2/12 had a persistent lesion. Those lesions were treated with electrocautery, and reached complete response in the following the 6 months (total of 18 months). In particular, there were no metachronous lesions detected. Patients reported moderate to severe pain during the first 24 h after RFA, but only mild discomfort was present at 30 days. Patients were asymptomatic at their 6- and 12-month visits. RFA was not associated with changes in anorectal manometry or ultrasound examination. The 36-SF survey reported improvement in the general health domain (p = 0.038), while the MGH-SFQ showed improvements in sexual function. CONCLUSIONS: In this study, targeted plus circumferential RFA had a 58.3% efficacy rate for the treatment of anal HSIL in immunocompromised patients, achieving 100% eradication after adding electrocautery ablation. No metachronous lesions were detected. Patients presented relatively mild symptoms after the procedure, no changes in anorectal anatomy or function, and some improvements in their sexual function. These results seem promising in light of the high recurrence reported after HSIL treatment. Larger studies are needed to validate our results.
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Neoplasias del Ano , Infecciones por Papillomavirus , Ablación por Radiofrecuencia , Lesiones Intraepiteliales Escamosas , Adulto , Neoplasias del Ano/cirugía , Femenino , Humanos , Masculino , Recurrencia Local de Neoplasia , Estudios RetrospectivosRESUMEN
BACKGROUND: The repressor element 1-silencing transcription factor (REST) is a regulator of gene expression, and the Ras association domain family member 1 A (RASSF1A) is an important tumor suppressor gene involved in cancer development. Although extensive characterization of the roles of REST and RASSF1A in cancer development have been reported in cellular models, the link between them and their possible role in the development of squamous intraepithelial lesions (SIL) and squamous cell carcinoma (SCC) of the cervix have not been explored. The aim of this study was to evaluate the expression of REST and RASSF1A in cervical cytological and histological samples from patients diagnosed with SIL or SCC and in CC-derived cell lines. METHODS: We analyzed the expression of REST and RASSF1A by immunocyto/histochemistry in cervical samples from patients (n = 271) and in cancer cell lines. Data analyses were performed using the Kruskal-Wallis test and generalized linear models. RESULTS: We identified binding sites for REST in RASSF1A and observed a significant reduction in REST and RASSF1A nuclear expression in samples from patients with high-grade SIL (HSIL) and SCC. For REST, we observed an average decrease of 334 and 423 r.u.d. for HSIL (n = 21) and SCC (n = 18) compared with non-LSIL (n = 72), whereas for RASSF1A, this decrease was 126 and 217 r.u.d., respectively (p < 0.001). CONCLUSION: Our results provide evidence of the altered expression of REST and RASSF1A in SIL and SCC, with a significant decrease in HSIL, SCC, and SCC-derived cell lines; findings that can contribute to the diagnosis, prognosis, and post-treatment follow-up of patients diagnosed with SIL or SCC.
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Carcinoma de Células Escamosas , Proteínas Represoras/genética , Lesiones Intraepiteliales Escamosas , Displasia del Cuello del Útero , Neoplasias del Cuello Uterino , Carcinoma de Células Escamosas/genética , Femenino , Humanos , Factores de Transcripción , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/genética , Neoplasias del Cuello Uterino/patología , Displasia del Cuello del Útero/diagnóstico , Displasia del Cuello del Útero/genética , Displasia del Cuello del Útero/patologíaRESUMEN
BACKGROUND/AIM: To determine the differential protein profiles of cervical cancer cell lines in order to find potential targets that can be used as biomarkers in low-grade squamous intraepithelial lesions (LSIL) diagnosis. MATERIALS AND METHODS: Proteomic analysis was performed on cervical cancer cell lines by 2D electrophoresis and liquid chromatography-mass spectrometry. Biomarker validation was performed in histological samples by immunofluorescence. RESULTS: Aldo-keto reductase C1 (AKR1C1) and transketolase-like 1 (TKTL1) proteins were selected as biomarkers and their expression was increased in samples with LSIL diagnosis. TKTL1 in combination with AKR1C1 increased sensitivity and specificity to 75% and 66%, respectively, with an area under curve of 0.76 in receiver operating characteristics curve analysis. CONCLUSION: AKR1C1 and TKTL1 showed potential as biomarkers for diagnosis of LSIL in Mexican women, with similar sensitivity and specificity to the biomarkers used in clinical trials for diagnosis of LSIL.
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20-Hidroxiesteroide Deshidrogenasas/metabolismo , Lesiones Intraepiteliales Escamosas/diagnóstico , Lesiones Intraepiteliales Escamosas/metabolismo , Transcetolasa/metabolismo , Displasia del Cuello del Útero/diagnóstico , Displasia del Cuello del Útero/metabolismo , Adulto , Área Bajo la Curva , Línea Celular Tumoral , Femenino , Humanos , México , Clasificación del Tumor , Proteínas de Neoplasias/metabolismo , Mapas de Interacción de Proteínas , Curva ROC , Lesiones Intraepiteliales Escamosas/patología , Displasia del Cuello del Útero/patologíaRESUMEN
Studies of cervical cancer (CC) have reported that microRNA-16-1 (miR-16-1), which is an oncomiR, is increased in the tissues and cell lines of CC. The aim of the present study was to investigate the association of miRNA-16-1 expression level with squamous cell carcinoma (SCC), the presence of squamous intraepithelial lesions (SIL) and the integration of high-risk human papillomavirus (HR-HPV) DNA. The current study analyzed 80 samples obtained from women by liquid-based cytology, which revealed that 20 were negative for SIL (NSIL) and without HPV, 20 were low-grade SIL (LSIL), 20 were high-grade SIL (HSIL), and 20 were diagnosed as SCC with HR-HPV. The genotyping of the viral DNA was conducted via an INNO-LiPA-HPV array, the expression of miR-16-1 was determined by reverse transcription-quantitative PCR, and the physical state of the HR-HPV was ascertained by in situ hybridization with amplification with tyramide. A total of eight HR-HPV genotypes were distinguished; the most frequent of these being HPV16, followed by multiple infection with HR-HPV (including HPV16). The mixed state of the HR-HPV was observed in 60 and 65% of LSIL and HSIL cases, respectively, while an integrated HR-HPV state was identified in 90% of cases with SCC. The expression level of miR-16-1 increased according to the grade of SIL, and cases with HSIL exhibited a significantly higher miR-16-1 expression level compared with women with NSIL (P<0.001; Table II). It can therefore be determined that the expression of miR-16-1 effects cellular proliferation, due to the viral integration of various HR-HPV genotypes in unique infection or in multiple infection. Thus, the overexpression of miR-16-1 could be monitored in women with LSIL, in order to discard a major lesion.
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PURPOSE: FOXP3 is a marker of the T regulatory (Treg) cell subset and drives its function and homeostasis. Its expression maintains the host immunosuppressive state that favors persistence of human papillomavirus (HPV) infection and squamous intraepithelial lesion (SIL) appearance. The present study evaluated the effects of the rs3761548 and rs2232365 intronic single-nucleotide variants (SNVs) and their haplotypes on HPV infection and SIL diagnosis in HPV-infected and -uninfected women. METHODS: HPV DNA-based detection in cervical specimens was performed by PCR. FOXP3 variants were genotyped by PCR-restriction fragment length polymorphism and haplotype recombination sites were inferred for 208 HPV-infected and 218 HPV-uninfected women diagnosed or not with low- or high-grade intraepithelial lesions of cervix. Case-control analyses were carried out by logistic regression adjusted for several socio-demographic, sexual lifestyle, and clinical data. RESULTS: The homozygous genotype of the rs3761548 variants (A/A) (related to decreased FOXP3 expression) may exert a protective role against HPV infection in women (ORAj: 0.60; 95% CI 0.36-0.99; p = 0.049) and was an independent predictor of protection against HSIL development (ORAdj: 0.28; 95% CI 0.11-0.68; p = 0.006). In addition, the homozygous genotype (G/G) of the rs2232365 variants (related to increased FOXP3 expression) was independently associated with the HPV infection (ORAdj: 2.10; 95% CI 1.06-4.15; p = 0.033). Haplotype analysis revealed no significant associations in our study. CONCLUSIONS: Our results reveal the significant and independent associations between FOXP3 genetic variants and susceptibility to HPV infection and SIL diagnosis and their role as biomarkers of HPV infection and cervical lesion management.
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Factores de Transcripción Forkhead/genética , Factores Inmunológicos/genética , Infecciones por Papillomavirus/diagnóstico , Polimorfismo de Nucleótido Simple , Lesiones Intraepiteliales Escamosas de Cuello Uterino/diagnóstico , Adulto , Brasil , Estudios de Casos y Controles , Estudios de Cohortes , Femenino , Predisposición Genética a la Enfermedad , Genotipo , Haplotipos , Humanos , Persona de Mediana Edad , Papillomaviridae/inmunología , Infecciones por Papillomavirus/genética , Reacción en Cadena de la Polimerasa , Polimorfismo de Longitud del Fragmento de Restricción , Pronóstico , Factores de Riesgo , Lesiones Intraepiteliales Escamosas de Cuello Uterino/genética , Neoplasias del Cuello Uterino/diagnóstico , Neoplasias del Cuello Uterino/genética , Adulto Joven , Displasia del Cuello del Útero/diagnóstico , Displasia del Cuello del Útero/genéticaRESUMEN
High-risk human papillomavirus (HPV) is the primary cause of cervical carcinoma (CC). Viral integration into the host chromosomes is associated with neoplastic progression, and epigenetic changes may occur as a result. The objective of the present study was to analyze HPV L1 gene methylation and to compare the use of quantitative polymerase chain reaction (qPCR), in situ hybridization (ISH) and L1 methylation analysis as methods for detecting HPV integration. Cervical scrapes or biopsy samples positive for HPV 16 or 18, from 187 female patients with CC, squamous intraepithelial lesions (SILs) or no intraepithelial lesion (non-IL) were analyzed. Methylation of the L1 gene was determined using bisulfite modification followed by PCR, and HPV integration was subsequently analyzed. HPV 16 L1 gene methylation was revealed to increase with histological grade, with statistically significant differences observed as follows: Low-grade SIL vs. CC, P<0.0001 and non-IL vs. CC, P<0.0001. HPV 18 L1 gene methylation also increased according to histological grade, however, no statistically significant differences were observed. Methylation at CpG site 5608 of the HPV 16 L1 gene was associated with all grades of cervical lesions, whereas methylation at CpG site 5617 demonstrated the strongest association with CC (odds ratio, 42.5; 95% confidence interval, 4.7-1861; P<0.0001). The concordance rates between the various methods for the detection of the physical status of HPV 16 and HPV 18 were 96.1% for qPCR and ISH, 76.7% for qPCR and L1 gene methylation, and 84.8% for ISH and L1 gene methylation. In conclusion, methylation of the HPV 16 L1 gene increases significantly according to the grade of the cervical lesion, and methylation at CpG sites 5608 and 5617 of this gene may be used as prognostic biomarkers. ISH and L1 gene methylation have good concordance with qPCR with regards to the detection of HPV integration. Therefore, these are useful methods in determining the physical state of HPV.
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Abstract Objective A vulvar squamous intraepithelial lesion is deemed to be a preceding lesion to vulvar cancer, especially in women aged under 40 years, holders of an acquired or idiopathic immunosuppression. Several treatments have been used to treat these lesions. One of the aesthetically acceptable therapeutic methods is the CO2 laser vaporization. Methods In a transversal study, 46 records of immunosuppressed women bearing a vulvar low grade and/or high grade squamous intraepithelial lesion were selected out of the retrospective analysis, computing age, date of record, date of vulvar lesion treatment with CO2 laser, the time elapsed between the first and the last visit (in months), the number of visits, the presence or absence of condylomatous lesions in other female lower genital tract sites and whether or not recurrences and persistence of intraepithelial lesions have been noticed during the follow-up. Results Patients bearing vulvar high-grade squamous intraepithelial lesion and immunosuppressed (serumpositive forhumanimmunodeficiency virus [HIV] or with solid organs transplantation) have shown a higher level of persistence of lesions and a higher chance of having other areas of the female lower genital tract involved. Conclusion While the CO2 laser vaporization is the most conservative method for the treatment of vulvar high-grade intraepithelial lesions, it is far frombeing the ideal method, dueto the intrinsic infection features considered. The possibility of persistence, recurrences and spontaneous limited regression indicates that a closer surveillance in the long-term treated cases should be considered, in special for immunosuppressed patients.
Resumo Objetivo A lesão intraepitelial escamosa da vulva é considerada uma lesão precursora do câncer vulvar, emespecial emmulheres com idade inferior a 40 anos, portadoras de imunossupressão adquirida ou idiopática. Vários tratamentos têm sido utilizados para tratar esse tipo de lesão. Um dos métodos terapêuticos esteticamente aceitáveis é a vaporização a laser CO2. Métodos Em um estudo transversal, foram selecionados da análise retrospectiva de prontuários arquivados no setor de patologia do trato genital inferior 46 prontuários de mulheres comimunossupressão e portadoras de lesão intraepitelial escamosa de baixo grau e/ou alto grau computando-se: idade, data de registro, data do tratamento da lesão vulvar com laser CO2, tempo entre a primeira e a última consulta (em meses), número de consultas, presença ou ausência de lesões condilomatosas em outros locais do aparelho reprodutor feminino e a ocorrência ou não de recidivas e persistência de lesões intraepiteliais durante o período de acompanhamento. Resultados Pacientes com lesão intraepitelial de alto grau vulvar e imunocomprometidas (soropositivas para HIV ou com transplante de órgãos sólidos) mostraram maior índice de persistência das lesões e maior chance de ter outras áreas do órgão genital feminino comprometidas. Conclusão Embora a vaporização a laser CO2 seja o método mais conservador para o tratamento das lesões intraepiteliais vulvares de alto grau, está longe de ser o método ideal pelas características intrínsecas da afecção em foco. A possibilidade de persistência, recidivas e regressão espontânea limitada recomenda maior vigilância para os casos tratados a longo prazo, em especial para as pacientes com imunossupressão.
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Humanos , Femenino , Adolescente , Adulto , Adulto Joven , Enfermedades de la Vulva/cirugía , Terapia por Láser , Láseres de Gas/uso terapéutico , Lesiones Intraepiteliales Escamosas de Cuello Uterino/sangre , Recurrencia , Estudios Transversales , Huésped Inmunocomprometido , Persona de Mediana EdadRESUMEN
Immunotherapy with dendritic cells (DCs) is a great promise for the treatment of neoplasms. However, the obtainment and protocol of differentiation of these cells may depend on extrinsic factors such as the tumor itself. The aim of the present study was to verify the influence of cervical neoplasia on different protocols of differentiation of monocyte-derived DCs resulting in an increased maturation phenotype. A total of 83 women were included in the study. The patients were grouped in low-grade squamous intraepithelial lesion (LSIL) (n=30), high-grade squamous intraepithelial lesion (HSIL) (n=22), cervical cancer (n=10) and healthy patients (n=21) groups. The mononuclear cells of patients were subjected to three differentiation protocols. In protocol I (pI), granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-4 and tumor necrosis factor (TNF)-α were used for the differentiation of mature DCs (pIDCs). In protocol II (pII), monocytes were stimulated with GM-CSF, IL-4, TNF-α and activated lymphocytes in the absence of non-adherent cells (pIIDCs). In protocol III (pIII), monocytes were stimulated with GM-CSF, IL-4, TNF-α and activated lymphocytes in the presence of non-adherent cells (pIIIDCs). These cells were evaluated by flow cytometry for the expression of maturation markers such as cluster of differentiation (CD)11c, CD86 and human leukocyte antigen-antigen D related (HLA-DR). The main cytokines secreted (IL-4, IL-12 and transforming growth factor-ß) were measured by ELISA. Our results indicate a significantly lower mature profile of pIIDCs and a significant increase in CD11c+ pIIIDCs able to produce IL-12 (P=0.0007). Furthermore, a significant reduction in cervical cancer HLA-DR+ pIDCs (P=0.0113) was also observed. HSIL patients exhibited a higher percentage of HLA-DR+ pIIDCs (P=0.0113), while LSIL patients had a lower percentage of CD11c+ pIIIDCs (P=0.0411). These findings suggest that the extent of cervical lesions affects the process of differentiation of DCs. Furthermore, activated lymphocytes may induce a better maturation of monocyte-derived DCs, and the presence of mononuclear cells appears to contribute to the DC differentiation process.
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OBJECTIVE: Atypical squamous cells of undetermined significance (ASCUS) and atypical glandular cells (AGC) reflect cellular abnormalities insufficient for clear diagnosis. We used cell pellets obtained from liquid-based cytology (LBC) to prepare cell blocks (CB) and clarify the initial diagnosis of ASCUS and AGC. STUDY DESIGN: A total of 393 CBs with initial diagnosis of ASCUS or AGC were processed. Of those, 305 of the ASCUS and 20 of the AGC had adequate specimens. We compared results of CBs prepared from ASCUS and AGC to determine which had higher frequencies of higher-grade lesions. RESULTS: A majority of specimens (83%) were adequate for evaluation. Compared with the initial diagnosis, 14% (42/305) of ASCUS were diagnosed with low-grade squamous intraepithelial lesion (LSIL) in CB, while 10% (2/20) of AGC were diagnosed with LSIL or adenocarcinoma. No statistical relationship between the initial diagnosis of ASCUS and AGC and results of higher-grade lesions in CB is evident as determined by p value greater than 0.05 (p = 0.228). CONCLUSIONS: CBs prepared from Liqui-PREP cell pellets are, in most cases, assessable and can be useful as an adjunctive test to help clarify the initial diagnosis of ASCUS and AGC. Diagn. Cytopathol. 2017;45:520-525. © 2017 Wiley Periodicals, Inc.
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Células Escamosas Atípicas del Cuello del Útero/patología , Células Epiteliales/patología , Prueba de Papanicolaou/métodos , Adulto , Femenino , Humanos , Prueba de Papanicolaou/normasRESUMEN
El cáncer de cuello uterino (CCU) es la primera causa de muerte por cáncer en mujeres en países en vías de desarrollo. La infección persistente por el virus papiloma humano (VPH) es un factor necesario en lesiones preneoplásicas y CCU. La citología cervicovaginal es el método mayormente utilizado para detectar el CCU y su uso combinado con la de detección de ADN viral seis meses post-tratamiento aumenta la efectividad para identificar mujeres tratadas con riesgo de lesión residual/recidiva. El objetivo fue describir la frecuencia de VPH de alto riesgo (AR) en mujeres tratadas por lesión escamosa intraepitelial (SIL) que acudieron al Servicio de Patología Cervical del Hospital San Pablo de enero-diciembre/2014. Se realizó un estudio descriptivo de corte transverso, que incluyó 80 pacientes que acudieron al servicio para control post-tratamiento por SIL. Se utilizó Cobas 4800 HPV Test (Roche) para la detección individual de VPH-16 y 18, y un pool de 10 VPH-AR (31,33,35,39,45,51,52,56,58,59) y dos de probable alto riesgo (66,68). Se encontró infección viral en el 7,5% (6/80) de las pacientes tratadas; identificándose VPH-16 en 3/6 de los casos positivos. En Paraguay existe alta incidencia de lesiones pre-neoplásicas y CCU siendo un problema de salud pública. Los datos observados sugieren que la utilización de este sistema para la detección viral puede llevar a optimizar el seguimiento post-tratamiento y la identificación de VPH-16 y 18 podría contribuir a la selección de pacientes en mayor riesgo de desarrollar una lesión cervical que deben someterse a una vigilancia frecuente y meticulosa.
Cervical cancer is the leading cause of cancer death in women in developing country.Persistent infection with human papillomavirus (HPV) is a necessary factor in premalignantlesions and cervical cancer. The Pap smear is the method most commonly used to detect thecervical cancer and its combined with the detection of viral DNA six months post-treatment increases effectiveness to identify women treated in risk with residual/recurrent lesion. Theaim was to describe the frequency of high-risk HPV (HR) in treated women for squamousintraepithelial lesion (SIL) who attended the Cervical Pathology Service of Hospital San Pablofrom January to December/2014. It is cross sectional, descriptive study which included 80patients who attended the service for post-treatment control. The 4800 Cobas HPV Test(Roche) was used, which detects HPV-16 and 18, and a pool of 10 HR-HPV(31,33,35,39,45,51,52,56,58,59) and two "probable" high risk (66,68). Viral infection wasfound in 7.5% (6/80) of patients treated; identifying HPV-16 in 3/6 of positive cases. InParaguay there is a high incidence of pre-neoplastic lesions and cervical cáncer remains apublic health problem. The observed data suggest that the use of this system for viraldetection can lead to optimize the post-treatment monitoring and the identification of HPV-16and 18 could contribute to the selection of patients at increased risk of developing cervicalinjury should undergo to frequent and careful monitoring.