RESUMO
PURPOSE: To investigate the frequency of a founder mutation in NLRP7, L750V, in independent cohorts of Mexican patients with recurrent hydatidiform moles (RHMs). METHODS: Mutation analysis was performed by Sanger sequencing on DNA from 44 unrelated Mexican patients with RHMs and seven molar tissues from seven additional unrelated patients. RESULTS: L750V was present in homozygous or heterozygous state in 37 (86%) patients and was transmitted on the same haplotype to patients from different states of Mexico. We also identified a second founder mutation, c.2810+2T>G in eight (18.1%) patients, and a novel premature stop-codon mutation W653*. CONCLUSION: Our data confirm the strong founder effect for L750V, which appears to be the most common mutation in NLRP7. We also report on six healthy live births to five patients with biallelic NLRP7 mutations, two from spontaneous conceptions and four from donated ovum and discuss our recommendations for DNA testing and genetic counseling.
Assuntos
Proteínas Adaptadoras de Transdução de Sinal/genética , Efeito Fundador , Mola Hidatiforme/genética , Mutação , Feminino , Haplótipos , Heterozigoto , Humanos , Nascido Vivo , México , Polimorfismo de Nucleotídeo Único , GravidezRESUMO
BACKGROUND: MicroRNAs are small noncoding RNAs with important regulatory functions. Although well-studied in cancer, little is known about the role of microRNAs in premalignant disease. Complete hydatidiform moles are benign forms of gestational trophoblastic disease that progress to gestational trophoblastic neoplasia in up to 20% of cases; however, there is no well-established biomarker that can predict the development of gestational trophoblastic neoplasia. OBJECTIVE: This study aimed to investigate possible differences in microRNA expression between complete moles progressing to gestational trophoblastic neoplasia and those regressing after surgical evacuation. STUDY DESIGN: Total RNA was extracted from fresh frozen tissues from 39 complete moles collected at the time of uterine evacuation in Brazil. In the study, 39 cases achieved human chorionic gonadotropin normalization without further therapy, and 9 cases developed gestational trophoblastic neoplasia requiring chemotherapy. Total RNA was also extracted from 2 choriocarcinoma cell lines, JEG-3 and JAR, and an immortalized normal placenta cell line, 3A-subE. MicroRNA expression in all samples was quantified using microRNA sequencing. Hits from the sequencing data were validated using a quantitative probe-based assay. Significantly altered microRNAs were then subjected to target prediction and gene ontology analyses to search for alterations in key signaling pathways. Expression of potential microRNA targets was assessed by quantitative real-time polymerase chain reaction and western blot. Finally, potential prognostic protein biomarkers were validated in an independent set of formalin-fixed paraffin-embedded patient samples from the United States (15 complete moles progressing to gestational trophoblastic neoplasia and 12 that spontaneously regressed) using quantitative immunohistochemistry. RESULTS: In total, 462 microRNAs were identified in all samples at a threshold of <1 tag per million. MicroRNA sequencing revealed a distinct set of microRNAs associated with gestational trophoblastic neoplasia. Gene ontology analysis of the most altered transcripts showed that the leading pathway was related to response to ischemia (P<.001). Here, 2 of the top 3 most significantly altered microRNAs were mir-181b-5p (1.65-fold; adjusted P=.014) and mir-181d-5p (1.85-fold; adjusted P=.014), both of which have been shown to regulate expression of BCL2. By quantitative real-time polymerase chain reaction, BCL2 messenger RNA expression was significantly lower in the complete moles progressing to gestational trophoblastic neoplasia than the regressing complete moles (-4.69-fold; P=.018). Reduced expression of BCL2 was confirmed in tissue samples by western blot. Immunohistochemistry in the independent patient samples revealed significantly lower cytoplasmic expression of BCL2 in the villous trophoblasts from cases destined for progression to gestational trophoblastic neoplasia compared with those that regressed, both with respect to staining intensity (optic density 0.110±0.102 vs 0.212±0.036; P<.001) and to the percentage of positive cells (16%±28% vs 49.4%±28.05%; P=.003). CONCLUSION: Complete moles progressing to gestational trophoblastic neoplasia are associated with a distinct microRNA profile. miR-181 family members and BCL2 may be prognostic biomarkers for predicting gestational trophoblastic neoplasia risk.
Assuntos
Progressão da Doença , Mola Hidatiforme/genética , MicroRNAs/metabolismo , Proteínas Proto-Oncogênicas c-bcl-2/genética , Neoplasias Uterinas/genética , Adolescente , Adulto , Feminino , Marcadores Genéticos , Doença Trofoblástica Gestacional/genética , Doença Trofoblástica Gestacional/patologia , Sequenciamento de Nucleotídeos em Larga Escala , Humanos , Mola Hidatiforme/patologia , MicroRNAs/genética , Pessoa de Meia-Idade , Gravidez , Proteínas Proto-Oncogênicas c-bcl-2/metabolismo , Neoplasias Uterinas/patologia , Adulto JovemRESUMO
BACKGROUND: Distinguishing hydatidiform moles (HMs) from nonmolar specimens and the subclassification of HM are important because complete hydatidiform mole (CHM) is associated with an increased risk of development of gestational trophoblastic neoplasia. However, diagnosis based solely on morphology has poor inter-observer reproducibility. Recent studies have demonstrated that the use of p57KIP2 immunostaining improves diagnostic accuracy for CHM. OBJECTIVES: To evaluate the accuracy of p57KIP2 immunostaining compared with molecular genotyping for the diagnosis of CHM. SEARCH STRATEGY: Major databases were searched from inception to March 2017 using the terms 'hydatidiform mole', 'p57', and 'genotyping', with their variations, and the search limit for the relevant study design. SELECTION CRITERIA: Any cross-sectional study, case series, case-control study, cohort study, or clinical trial that evaluated the accuracy of p57KIP2 immunostaining for the diagnosis of CHM compared with genotyping was included. Case reports, narrative reviews, expert opinions, and animal testing were excluded. DATA COLLECTION AND ANALYSIS: Extracted accuracy data were tabulated and pooled using a hierarchical bivariate random effects model. MAIN RESULTS: Bivariate meta-analysis produced a summary sensitivity of 0.984 (95% CI: 0.916-1.000) and specificity of 0.625 (95% CI: 0.503-0.736) with significant heterogeneity for specificity (I2 = 71.8, chi-square P = 0.029). The pooled summary diagnostic odds ratio was 56.54 (95% CI: 11.03-289.74) with no heterogeneity (I2 = 0.00%, chi-square P = 0.67). The diagnostic performance of the test was high with an area under the curve of (AUC) 0.980. CONCLUSIONS: p57KIP2 immunostaining is accurate when diagnosing CHM. It can be used as an adjunct test in a combination algorithmic approach. TWEETABLE ABSTRACT: A meta-analysis to evaluate the accuracy of p57KIP2 compared with genotyping to diagnose CHM.
Assuntos
Inibidor de Quinase Dependente de Ciclina p57/genética , Genótipo , Mola Hidatiforme/diagnóstico , Neoplasias Uterinas/diagnóstico , Feminino , Humanos , Mola Hidatiforme/genética , Imuno-Histoquímica , Gravidez , Sensibilidade e Especificidade , Neoplasias Uterinas/genéticaRESUMO
BACKGROUND: Distinguishing hydatidiform moles (HMs) from non-molar specimens and the subclassification of HM are important because complete hydatidiform mole (CHM) is associated with an increased risk of gestational trophoblastic neoplasia. However, diagnosis based solely on morphology has poor interobserver reproducibility. Recent studies have demonstrated that the use of p57KIP2 immunostaining improves diagnostic accuracy for CHM. METHODS: We will conduct a systematic review of prospective and retrospective studies to evaluate the accuracy of p57KIP2 immunostaining compared with molecular genotyping for the diagnosis of CHM. A high-sensitivity search strategy will be employed in MEDLINE, EMBASE, LILACS, The Grey Literature Report, OpenGrey, OAIster, and Cochrane CENTRAL. Two reviewers will independently screen all identified references for eligibility and extract data. The methodological quality and bias of the included studies will be assessed according to the Quality Assessment of Diagnostic Accuracy Studies (QUADAS-2) tool, and the overall quality of evidence will be assessed using the Grading of Recommendations Assessment, Development, and Evaluation (GRADE) approach. If a meta-analysis is possible, pooled estimates of sensitivity, specificity, and positive and negative likelihood ratios will be calculated using bivariate random-effects models. Statistical heterogeneity will be evaluated with I 2 statistics and explored through sensitivity analysis. DISCUSSION: There is considerable overlap between the histological features of molar and non-molar pregnancies and between complete and partial HMs, which results in significant interobserver variability in the diagnosis of CHM and its mimics. Therefore, molecular techniques are used to correctly diagnosis and treat CHM. However, these molecular diagnostic methods are technically difficult to perform, relatively costly, and unavailable in most pathology laboratories. According to our results, p57KIP2 immunostaining appears to be a practical and accurate adjunct for the diagnosis of CHM and its mimics because this technique is relatively simple, reliable, cost-efficient, and rapid. This systematic review will help to determine whether p57KIP2 immunostaining is an adequate alternative diagnostic test for CHM. SYSTEMATIC REVIEW REGISTRATION: PROSPERO CRD42015024181.
Assuntos
Inibidor de Quinase Dependente de Ciclina p57/genética , Genótipo , Mola Hidatiforme/diagnóstico , Mola Hidatiforme/genética , Neoplasias Uterinas/genética , Testes Diagnósticos de Rotina/normas , Feminino , Humanos , Mola Hidatiforme/patologia , Imuno-Histoquímica/métodos , Reação em Cadeia da Polimerase , Gravidez , Reprodutibilidade dos Testes , Revisões Sistemáticas como Assunto , Neoplasias Uterinas/diagnóstico , Neoplasias Uterinas/patologiaRESUMO
BACKGROUND: TJP1 gene encodes a ZO-1 protein that is required for the recruitment of occludins and claudins in tight junction, and is involved in cell polarisation. It has different variations, the frequency of which has been studied in different populations. In Mexico there are no studies of this gene. These are required because their polymorphisms can be used in studies associated with medicine and surgery. Therefore, the aim of this study was to estimate the frequency of alleles and genotypes of rs2291166 gene polymorphism TJP1 in Mexico Mestizos population, and to estimate the conformational effect of an amino acid change. MATERIAL AND METHODS: A total of 473 individuals were included. The rs2291166 polymorphism was identified PASA PCR-7% PAGE, and stained with silver nitrate. The conformational effect of amino acid change was performed in silico, and was carried out with servers ProtPraram Tool and Search Database with Fasta. RESULTS: The most frequent allele in the two populations is the ancestral allele (T). A genotype distribution similar to other populations was found. The polymorphism is in Hardy-Weinberg, p>0.05. Changing aspartate to alanine produced a conformational change. CONCLUSIONS: The study reveals a high frequency of the ancestral allele at rs2291166 polymorphism in the Mexican population.
Assuntos
Etnicidade/genética , Polimorfismo de Nucleotídeo Único , Proteína da Zônula de Oclusão-1/genética , Adolescente , Adulto , Idoso , Idoso de 80 Anos ou mais , Alelos , Sequência de Aminoácidos , Substituição de Aminoácidos , Simulação por Computador , Feminino , Frequência do Gene , Predisposição Genética para Doença , Genótipo , Humanos , Mola Hidatiforme/genética , Indígenas Norte-Americanos/genética , Masculino , Casamento , México , Pessoa de Meia-Idade , Modelos Genéticos , Modelos Moleculares , Dados de Sequência Molecular , Síndromes Neoplásicas Hereditárias/genética , Pancreatite/genética , Gravidez , Conformação Proteica , Estabilidade Proteica , Espanha/etnologia , Adulto Jovem , Proteína da Zônula de Oclusão-1/químicaRESUMO
OBJECTIVE: The aim of this study is to analyze NLRP7 mutation frequency in 20 Mexican patients with recurrent hydatidiform moles (RHMs). PATIENTS: Twenty patients with RHMs, 50 couples with recurrent pregnancy loss (RPL), and 100 controls were included in the study. Molecular analysis of the NLRP7 coding region was performed in patients with RHMs. Restriction enzyme digestion analysis and direct sequencing of the identified mutations were performed in controls and patients with RPL. RESULTS: Patients displayed between two and six moles, and 10 of them presented other forms of pregnancy loss. Twelve (60%) patients were homozygous for the missense mutation c.2248C > G (p.L750V), five (25%) patients were heterozygous for the p.L750V mutation and the c.1018 G > A (p.E340K) variant, and three (15%) patients were heterozygous for the c.1018 G > A (p.E340K) variant. Five (5%) control women and four women and one man (5%) with RPL were heterozygous for the p.L750V mutation and two (2%) patients with RPL were heterozygous for the p.E340K variant. CONCLUSIONS: A total of 60% of our RHM patients presented homozygous p.L750V mutations, 25% were compound heterozygotes for p.L750V mutation and the p.E340K variant, and 15% were heterozygous for p.E340K variant. Heterozygous p.L750V mutations were frequently observed in our population. Homozygous mutations were also present in patients with RHMs. Additional studies are needed to understand the role of the p.E340K variant in RHMs and RPL.
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Proteínas Adaptadoras de Transdução de Sinal/genética , Mola Hidatiforme/genética , Neoplasias Uterinas/genética , Aborto Habitual/etnologia , Aborto Habitual/genética , Adulto , Estudos de Casos e Controles , Feminino , Predisposição Genética para Doença , Heterozigoto , Homozigoto , Humanos , Mola Hidatiforme/etnologia , México , Mutação , Taxa de Mutação , Mutação de Sentido Incorreto/genética , Recidiva Local de Neoplasia , Gravidez , Neoplasias Uterinas/etnologia , Adulto JovemRESUMO
A mola hidatiforme (MH) é a forma mais comum de doença trofoblástica gestacional e representa uma condição benigna que em alguns casos pode sofrer malignização. Todas as pacientes diagnosticadas com doenças molares são acompanhadas por pelo menos seis meses para detecção precoce da neoplasia trofoblástica gestacional. No momento, existem poucas ferramentas para avaliação prognóstica da mola hidatiforme. Foi descrita a expressão diferencial de diversos fatores em tecido molar em comparação ao trofoblasto não neoplásico. Essas moléculas podem estar relacionadas com o comportamento agressivo da MH e consequentemente poderiam servir para melhor entendimento do processo de malignização e como preditoras da evolução da doença trofoblástica gestacional.
The hydatidiform mole (HM) is the most common form of gestational trophoblastic disease and a benign condition that in some cases may undergo malignant transformation. All patients diagnosed with molar diseases are monitored for at least six months for early detection of gestational trophoblastic neoplasia. Currently, there are few prognostic tools for the prediction of hydatidiform mole evolution. Differential expression on molar tissue of different molecular factors have been described when compared to non-neoplastic trophoblast. These markers may be associated with aggressive behavior of HM and therefore could serve as predictors of the development of gestational trophoblastic disease and to better understand molar malignant transformation. This review article will summarize and evaluate prognostic molecular markers of HM.
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Humanos , Masculino , Feminino , Expressão Gênica , Mola Hidatiforme/etiologia , Mola Hidatiforme/genética , Transformação Celular Neoplásica , Progressão da Doença , Doença Trofoblástica Gestacional/genética , Detecção Precoce de Câncer , Imuno-Histoquímica , Biomarcadores Tumorais/análise , Regressão Neoplásica Espontânea , PrognósticoRESUMO
The StAR-related lipid transfer (START) domain is defined as a motif of around 200 amino acids implicated in lipid/sterol binding. In a previous study, we identified the StarD7 transcript encoding one of the 15 family members with START domain present in the human genome. This transcript was found to be overexpressed in choriocarcinoma JEG-3 cells. In addition, we demonstrated that the recombinant StarD7 protein forms stable Gibbs and Langmuir monolayers at the air-buffer interface, showing marked surface activity and interaction with phospholipid monolayers, mainly with phosphatidylserine, cholesterol and phosphatidylglycerol. This study was undertaken to evaluate the expression and localization of StarD7 protein in trophoblastic samples. Here, we show for the first time the presence of StarD7 protein in human trophoblast cells. Western blot assays revealed a unique specific 34 kDa protein in JEG-3 cell line, choriocarcinoma tissue, complete hydatidiform mole, early and normal term placenta. Immunohistochemical data from early and normal term placentas and complete hydatidiform moles showed that this protein is abundant in the syncytiotrophoblasts, mainly at the apical side of the syncytium, with a weak and focal reaction in the cytotrophoblast cells. Furthermore, an increased StarD7 mRNA and protein expression, as well as a change in its sub-cellular localization was observed in in vitro differentiating cytotrophoblast isolated from normal term placenta. Taken together, these findings support and allow future studies to explore the possibility that StarD7 protein mediates transplacental lipid transport and/or is involved in syncytialization.
Assuntos
Proteínas de Transporte/genética , Proteínas de Transporte/metabolismo , Trofoblastos/metabolismo , Animais , Células COS , Diferenciação Celular/genética , Células Cultivadas , Chlorocebus aethiops , Feminino , Humanos , Mola Hidatiforme/genética , Mola Hidatiforme/metabolismo , Gravidez , Nascimento a Termo/metabolismo , Distribuição Tecidual , Neoplasias Uterinas/genética , Neoplasias Uterinas/metabolismoRESUMO
OBJECTIVE: To explore the utility of histological, immunohistochemical and chromosome in situ hybridization (CISH) test in the differential diagnosis of Complete Hydatidiform Mole (CHM), Partial Hydatidiform Mole (PHM) and Hydropic Abortion (HA). STUDY DESIGN: We analyzed the histological characteristics, p57kip2 and Factor VIII expression and CISH test in 38 cases with some diagnostic concerns, comprising 13 CHM, 14 PHM and 11 HA. RESULTS: Our results indicate that p57kip2 expression and the ploidy assessed by CISH were essential for the reclassification of 2 cases, one from CHM to PHM and another from PHM to HA, as well as for confirming the previous diagnosis in cases where there were conflicting features. p57kip2 expression is diagnostic if no cells at all present it (CHM) or when there are over 10% of cells expressing it (PHM and HA). CONCLUSIONS: We concluded that there is no single criterion for the distinction of CHM, PHM and HA. So p57kip2 expression and CISH test can be used in association with the histological findings for the differential diagnosis of the three conditions in cases presenting some concern for definitive diagnosis.
Assuntos
Aborto Espontâneo/diagnóstico , Mola Hidatiforme/diagnóstico , Neoplasias Uterinas/diagnóstico , Aborto Espontâneo/genética , Aborto Espontâneo/metabolismo , Adolescente , Adulto , Inibidor de Quinase Dependente de Ciclina p57/metabolismo , Diagnóstico Diferencial , Fator VIII/metabolismo , Feminino , Humanos , Mola Hidatiforme/genética , Mola Hidatiforme/metabolismo , Imuno-Histoquímica , Hibridização In Situ , Gravidez , Estudos Retrospectivos , Neoplasias Uterinas/genética , Neoplasias Uterinas/metabolismoRESUMO
AIM: Considering the roles of c-erB-2 and p53 oncoproteins in tumor progression, we aimed to evaluate their expression in hydatidiform moles, and the possible predictive value of this immunoexpression in postmolar follow-up. METHODS: Group I comprised 35 patients with progression to gestational trophoblastic tumor, and group II included 32 patients with progression to spontaneous remission. Immunohistochemical tests were performed by streptavidin-peroxidase method. c-erbB-2 immunoexpression was evaluated according to quantitative and semiquantitative criteria; p53 according to percentage of cells with stained nuclei. Data were analyzed by Student t-test, Mann-Whitney test, ROC curve and logistic regression analysis. RESULTS: c-erbB-2 and p-53 expressions were significantly increased in group I. Quantitative and semiquantitative analysis of c-erb-2 showed that its expression may be associated with mole hydatidiform progression to gestational trophoblastic tumor. Taking into account cells with complete membranous delineation we proposed a cut-off value of 10.8%. Similarly, considering the percentage of cells presenting nuclei marked by p53 we suggested a cut-off value of 40.1% for the prediction of malignant transformation of mole hydatidiform. CONCLUSIONS: c-erbB-2 and p53 immunoexpression in hydatidiform mole are usually increased with malignant transformation. In addition to beta-fraction of human chorionic gonadotropin, they could possibly help the establishment of a therapeutic protocol.