RESUMO
OBJECTIVE: Our study aimed to identify mutations in the FMR1 gene in a group of Brazilian women diagnosed with primary ovarian insufficiency (POI). METHODS: This cross-sectional study included patients aged under 40 years with confirmed POI from a convenience sample of patients seen from June 2017 to December 2018 at a University Hospital in Curitiba, Brazil. Genomic DNA was extracted and analyzed using FragilEase(tm) PCR kits (PerkinElmer), a commercially available test that enables the quantification of CGG trinucleotide repeat expansions in the FMR1 gene. RESULTS: A total of 52 patients with an average age of 35.8±3.97 years were included. Fifty (96.1%) had normal alleles with 18 to 43 CGG repeats. The most frequent CGG-repeat sizes were 28 and 30. Two patients (3.8%) presented mutations in the FMR1 gene. The first had alleles with 19/97 CGG repeats, was categorized as a premutation carrier for FXS, and had a son with cognitive impairment. The second had alleles with 21/45 CGG repeats and was described as belonging to the gray zone. CONCLUSIONS: In our study, 3.8% of the females with POI had mutations in the FMR1 gene. The most frequent allele sizes were 28 and 30 CGG repeats.
Assuntos
Proteína do X Frágil da Deficiência Intelectual , Doenças Ovarianas , Insuficiência Ovariana Primária , Adulto , Brasil/epidemiologia , Estudos Transversais , Feminino , Proteína do X Frágil da Deficiência Intelectual/genética , Humanos , Mutação , Doenças Ovarianas/genética , Insuficiência Ovariana Primária/genética , Repetições de TrinucleotídeosRESUMO
OBJECTIVE: Ovarian endometriosis seriously affects the quality of life of females, and long non-coding RNA lncRNA urothelial carcinoma-associated 1 (UCA1) plays pivotal roles in the pathogenesis of various ovarian diseases. However, the involvement of lncRNA UCA1 in ovarian endometriosis remains unknown to date. Therefore, the present study aims to study the role of UCA1 in ovarian endometriosis. METHODS: A total of 98 patients with ovarian endometriosis and 28 healthy females were included. The expression of lncRNA UCA1 in ectopic and eutopic endometrium tissues of ovarian endometriosis patients and controls was detected using qRT-PCR. A ROC curve analysis was performed to evaluate the diagnostic values of serum lncRNA UCA1 for ovarian endometriosis. Patients were followed up for 2 years after discharge, and the recurrence of ovarian endometriosis was recorded. RESULTS: The expression level of lncRNA UCA1 was significantly higher in ectopic endometrium tissues than in paired eutopic endometrium tissues for most of the patients. The serum lncRNA UCA1 level showed no significant correlations with either patients' age or living habits. After the treatment, the serum lncRNA UCA1 level increased, and serum levels of lncRNA UCA1 on the day of discharge were significantly lower in patients with recurrence than those in patients without recurrence. Conclusion: The downregulation of lncRNA UCA1 is involved in the pathogenesis of ovarian endometriosis and may serve as a promising diagnostic and prognostic biomarker for the disease.
Assuntos
Regulação para Baixo , Endometriose/sangue , Endometriose/diagnóstico , Doenças Ovarianas/sangue , Doenças Ovarianas/diagnóstico , RNA Longo não Codificante/sangue , Adulto , Análise de Variância , Biomarcadores/sangue , Estudos de Casos e Controles , Endometriose/genética , Endométrio/patologia , Feminino , Humanos , Doenças Ovarianas/genética , Prognóstico , RNA Longo não Codificante/genética , Reação em Cadeia da Polimerase em Tempo Real , Recidiva , Valores de Referência , Sensibilidade e Especificidade , Adulto JovemRESUMO
SUMMARY OBJECTIVE: Ovarian endometriosis seriously affects the quality of life of females, and long non-coding RNA lncRNA urothelial carcinoma-associated 1 (UCA1) plays pivotal roles in the pathogenesis of various ovarian diseases. However, the involvement of lncRNA UCA1 in ovarian endometriosis remains unknown to date. Therefore, the present study aims to study the role of UCA1 in ovarian endometriosis. METHODS: A total of 98 patients with ovarian endometriosis and 28 healthy females were included. The expression of lncRNA UCA1 in ectopic and eutopic endometrium tissues of ovarian endometriosis patients and controls was detected using qRT-PCR. A ROC curve analysis was performed to evaluate the diagnostic values of serum lncRNA UCA1 for ovarian endometriosis. Patients were followed up for 2 years after discharge, and the recurrence of ovarian endometriosis was recorded. RESULTS: The expression level of lncRNA UCA1 was significantly higher in ectopic endometrium tissues than in paired eutopic endometrium tissues for most of the patients. The serum lncRNA UCA1 level showed no significant correlations with either patients' age or living habits. After the treatment, the serum lncRNA UCA1 level increased, and serum levels of lncRNA UCA1 on the day of discharge were significantly lower in patients with recurrence than those in patients without recurrence. Conclusion: The downregulation of lncRNA UCA1 is involved in the pathogenesis of ovarian endometriosis and may serve as a promising diagnostic and prognostic biomarker for the disease.
RESUMO OBJETIVO: A endometriose ovariana afeta seriamente a qualidade de vida das mulheres, e o carcinoma urotelial 1 de urcélio de RNA não codificador longo 1 (UCA1) desempenha um papel crucial na patogênese de várias doenças ovarianas. No entanto, o envolvimento do lncRNA UCA1 na endometriose ovariana permanece desconhecido até o momento. Portanto, o presente estudo tem como objetivo estudar o papel do UCA1 na endometriose ovariana. Métodos: Um total de 98 pacientes com endometriose ovariana e de 28 mulheres saudáveis foi incluído. A expressão de lncRNA UCA1 em tecidos de endométrio ectópico e eutópico de pacientes com endometriose ovariana e controles foi detectada por qRT-PCR. A análise da curva ROC foi realizada para avaliar os valores diagnósticos do lncRNA UCA1 sérico para endometriose ovariana. Os pacientes foram acompanhados por dois anos após a alta, e a recorrência da endometriose ovariana foi registrada. RESULTADOS: O nível de expressão do lncRNA O UCA1 foi significativamente maior nos tecidos do endométrio ectópico do que nos tecidos do endométrio eutópico pareados para a maioria dos pacientes. O nível sérico de UCA1 foi diminuído com a progressão da endometriose ovariana. O soro UCA1 pode ser usado para diagnosticar com precisão a endometriose ovariana. O nível sérico de UCA1 não apresentou correlações significativas com a idade ou com os hábitos de vida dos pacientes. Após o tratamento, o nível sérico do lncRNA UCA1 foi aumentado, e os níveis séricos de lncRNA UCA1 no dia da alta foram significativamente menores nos pacientes com recidiva do que naqueles sem recorrência. CONCLUSÃO: A regulação negativa do lncRNA UCA1 está envolvida na patogênese da endometriose ovariana e pode servir como um promissor biomarcador diagnóstico e prognóstico para a doença.
Assuntos
Humanos , Feminino , Adulto , Adulto Jovem , Doenças Ovarianas/diagnóstico , Doenças Ovarianas/sangue , Regulação para Baixo , Endometriose/diagnóstico , Endometriose/sangue , RNA Longo não Codificante/sangue , Doenças Ovarianas/genética , Recidiva , Valores de Referência , Biomarcadores/sangue , Estudos de Casos e Controles , Prognóstico Clínico Dinâmico Homeopático , Análise de Variância , Sensibilidade e Especificidade , Endometriose/genética , Endométrio/patologia , Reação em Cadeia da Polimerase em Tempo Real , RNA Longo não Codificante/genéticaRESUMO
Endometriosis is a benign, estrogen-dependent disease with symptoms such as pelvic pain and infertility, and it is characterized by the ectopic distribution of endometrial tissue. The expression of the ID2, PRELP and SMOC2 genes was compared between the endometrium of women without endometriosis in the proliferative phase of their menstrual cycle and the eutopic and ectopic endometrium of women with endometriosis in the proliferative phase. Paired tissue samples from 20 women were analyzed: 10 from endometrial and peritoneal endometriotic lesions and 10 from endometrial and ovarian endometriotic lesions. As controls, 16 endometrium samples were collected from women without endometriosis in the proliferative phase of menstrual cycle. Analysis was performed by real-time polymerase chain reaction (PCR). There was no significant difference between gene expression in the endometrium of women with and without endometriosis. The ID2 gene expression was increased in the most advanced stage of endometriosis and in ovarian endometriomas, the PRELP was more expressed in peritoneal lesions, and the SMOC2 was highly expressed in both peritoneal and endometrioma lesions. Considering that the genes studied participate either directly or indirectly in cellular processes that can lead to cell migration, angiogenesis, and inappropriate invasion, it is possible that the deregulation of these genes caused the development and maintenance of ectopic tissue.
Assuntos
Endometriose/genética , Proteínas da Matriz Extracelular/genética , Glicoproteínas/genética , Proteína 2 Inibidora de Diferenciação/genética , Osteonectina/genética , Doenças Ovarianas/genética , Doenças Peritoneais/genética , Adolescente , Adulto , Estudos de Casos e Controles , Endometriose/metabolismo , Proteínas da Matriz Extracelular/metabolismo , Feminino , Regulação da Expressão Gênica , Glicoproteínas/metabolismo , Humanos , Proteína 2 Inibidora de Diferenciação/metabolismo , Ciclo Menstrual , Osteonectina/metabolismo , Doenças Ovarianas/metabolismo , Doenças Peritoneais/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Adulto JovemRESUMO
OBJECTIVE: Endometriosis is a multifactorial gynecological disease, whose pathogenesis is crucially dependent on angiogenesis, which is signaled via vascular endothelial growth factor (VEGF) and its receptor (VEGFR2). We hypothesize that single nucleotide polymorphisms (SNPs) in VEGF and VEGFR2 genes may influence the onset and/or the progression of endometriosis. The main aim of this study was to investigate the contribution of VEGF and VEGFR2 SNPs as risk factors for endometriosis, as well as their association with endometriosis symptoms. STUDY DESIGN: A case-control study was conducted, involving 293 endometriosis patients and 223 controls, who were submitted to laparoscopic or laparotomy surgery at hospitals from the Brazilian public health system. Genotyping of VEGF (-2578C>A, -460T>C, -1154G>A, +405G>C and +936C>T) and VEGFR2 (-604T>C, 1192C>T) SNPs was performed by TaqMan real-time polymerase chain reaction. The association between SNPs and endometriosis, deep infiltrating endometriosis (DIE) or endometriosis symptoms was estimated by odds ratios (OR) with their 95% confidence intervals (CI), which were calculated using multivariate logistic regression models. RESULTS: VEGF variant alleles -2578A and -1154A were associated with increased endometriosis risk (OR: 1.39, 95% CI: 1.04-1.87 and OR: 1.63, 95% CI: 1.12-2.37, respectively), whereas VEGF 405C and VEGFR2 1192T were associated with lower risk of endometriosis (OR: 0.66, 95% CI: 0.43-1.00 and OR: 0.58, 95% CI: 0.40-0.84, respectively). The combination of wild-type genotypes of both VEGF -2578C>A and -1154G>A with variant genotypes of both VEGF +405G>C and VEGFR2 1192C>T showed the best protective effect against the development of endometriosis, either considering all cases (OR: 0.33, 95% CI: 0.12-0.89) or only DIE (OR: 0.30, 95% CI: 0.10-0.87). The combination of variant genotypes of VEGF -2578C>A, -1154G>A, +405G>C and VEGFR2 1192C>T was also protective against DIE (OR: 0.67, 95% CI: 0.46-0.96). VEGFR2 1192C>T were associated with reduced cyclical urinary complaints (OR: 0.40, 95% CI: 0.18-0.88). CONCLUSIONS: Our results indicate that VEGF SNPs -2578C>A and -1154G>A increase endometriosis risk, whereas VEGF +405G>C and VEGFR2 1192C>T are protective against disease development, with VEGFR2 1192C>T also reducing cyclical urinary symptoms. The combined analysis of VEGF-VEGFR2 genotypes suggests a gene-gene interaction in endometriosis susceptibility.
Assuntos
Endometriose/genética , Doenças Ovarianas/genética , Polimorfismo de Nucleotídeo Único , Receptores de Fatores de Crescimento do Endotélio Vascular/genética , Fator A de Crescimento do Endotélio Vascular/genética , Adulto , Estudos de Casos e Controles , Epistasia Genética , Feminino , Predisposição Genética para Doença , Genótipo , Humanos , Pessoa de Meia-Idade , Adulto JovemRESUMO
Endometriosis is an estrogen-dependent disease affecting up to 10% of all premenopausal women. There is evidence that different endometriosis sites show distinct local estrogen concentration, which, in turn, might be due to a unique local estrogen metabolism. We aimed to investigate whether there was a site-specific regulation of selected enzymes responsible for the oxidative metabolism of estrogens in biopsy samples and endometrial and endometriotic stromal cells. Cytochrome P450 (CYP) 1A1 and CYP1B1 mRNA and protein expressions in deep-infiltrating (rectal, retossigmoidal, and uterossacral) lesions, superficial (ovarian and peritoneal) lesions, and eutopic and healthy (control) endometrium were evaluated by real-time PCR and western blot. Using a cross-sectional study design with 58 premenopausal women who were not under hormonal treatment, we were able to identify an overall increased CYP1A1 and CYP1B1 mRNA expression in superficial lesions compared with the healthy endometrium. CYP1A1 mRNA expression in superficial lesions was also greater than in the eutopic endometrium. Interestingly, we found a similar pattern of CYP1A1 and CYP1B1 expression in in vitro stromal cells isolated from ovarian lesions (n=3) when compared with stromal cells isolated from either rectum lesions or eutopic endometrium. In contradiction, there was an increased half-life of estradiol (measured by HPLC-MS-MS) in ovarian endometriotic stromal cells compared with paired eutopic stromal endometrial cells. Our results indicate that there is a site-dependent regulation of CYP1A1 and CYP1B1 in ovarian/peritoneal lesions and ovarian endometriotic stromal cells, whereas a slower metabolism is taking place in these cells.
Assuntos
Citocromo P-450 CYP1A1/genética , Citocromo P-450 CYP1B1/genética , Endometriose/genética , Endométrio/metabolismo , Doenças Ovarianas/genética , Doenças Peritoneais/genética , Adulto , Estudos de Casos e Controles , Estudos Transversais , Citocromo P-450 CYP1A1/metabolismo , Citocromo P-450 CYP1B1/metabolismo , Endometriose/metabolismo , Feminino , Humanos , Doenças Ovarianas/metabolismo , Doenças Peritoneais/metabolismoRESUMO
OBJECTIVE: To perform a clinical, biochemical, and molecular evaluation of patients with CYP17A1 defects, including ovarian imaging. DESIGN: Retrospective study. SETTING: Tertiary care center. PATIENT(S): Sixteen patients with congenital adrenal hyperplasia due to CYP17A1 defects with a median chronological age of 20 years and belonging to 10 unrelated families. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Clinical and biochemical parameters, molecular diagnosis, ovarian imaging, and therapeutic management. RESULT(S): Seventy-one percent of patients presented with primary amenorrhea, 50% had no breast development, and pubic hair was absent or sparse in all patients; 88% had high blood pressure at diagnosis. Basal LH and P levels were high, and androgen levels were low in all patients. Ultrasound revealed ovarian enlargement in 68.7% and ovarian macrocysts in 62.5% of patients before treatment; three patients had a previous surgical correction of ovarian torsion or rupture. Molecular analysis revealed inactivating CYP17A1 mutations in all patients. The most prevalent mutation was p.W406R, and one patient bore a novel p.G478S/p.I223Nfs*10 compound heterozygous mutation. Treatment with dexamethasone, estrogen, and P resulted in reduction of ovarian volume. CONCLUSION(S): Amenorrhea, absent/sparse pubic hair, hypertension, and ovarian macrocysts, whichincrease the risk of ovarian torsion, are important elements in the diagnosis of 46,XX patients with CYP17A1 defects. High basal P levels in patients with hypergonadotropic hypogonadism point to the diagnosis of CYP17A1 defects. Fertility can be achieved in these patients with novel reproductive techniques.
Assuntos
Transtornos 46, XX do Desenvolvimento Sexual/genética , Corticosteroides , Hiperplasia Suprarrenal Congênita/genética , Doenças Ovarianas/genética , Esteroide 17-alfa-Hidroxilase/genética , Transtornos 46, XX do Desenvolvimento Sexual/sangue , Transtornos 46, XX do Desenvolvimento Sexual/diagnóstico , Adolescente , Corticosteroides/sangue , Hiperplasia Suprarrenal Congênita/sangue , Hiperplasia Suprarrenal Congênita/diagnóstico , Adulto , Criança , Feminino , Humanos , Doenças Ovarianas/sangue , Doenças Ovarianas/diagnóstico , Linhagem , Estudos Retrospectivos , Adulto JovemRESUMO
Oestrogens play an important role in development and function of the brain and reproductive tract. Accordingly, it is considered that developmental exposure to environmental oestrogens can disrupt neural and reproductive tract development, potentially resulting in long-term alterations in neurobehaviour and reproductive function. Many chemicals have been shown to have oestrogenic activity, whereas others affect oestrogen production and turnover, resulting in the disruption of oestrogen signalling pathways. However, these mechanisms and the concentrations required to induce these effects cannot account for the myriad adverse effects of environmental toxicants on oestrogen-sensitive target tissues. Hence, alternative mechanisms are assumed to underlie the adverse effects documented in experimental animal models and thus could be important to human health. In this review, the epigenetic regulation of gene expression is explored as a potential target of environmental toxicants including oestrogenic chemicals. We suggest that toxicant-induced changes in epigenetic signatures are important mechanisms underlying the disruption of ovarian follicular development. In addition, we discuss how exposure to environmental oestrogens during early life can alter gene expression through effects on epigenetic control potentially leading to permanent changes in ovarian physiology.
Assuntos
Poluentes Ambientais/toxicidade , Epigênese Genética/efeitos dos fármacos , Estrogênios/toxicidade , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Doenças Ovarianas/induzido quimicamente , Doenças Ovarianas/fisiopatologia , Ovário/efeitos dos fármacos , Ovário/fisiopatologia , Animais , Epigênese Genética/genética , Feminino , Regulação da Expressão Gênica no Desenvolvimento/genética , Humanos , Doenças Ovarianas/genética , Ovário/crescimento & desenvolvimentoRESUMO
Considerable effort has been invested in searching for less invasive methods of diagnosing endometriosis. Previous studies have indicated altered levels of the CALD1 gene (encoding the protein caldesmon) in endometriosis. The aims of our study were to investigate whether average CALD1 expression and caldesmon protein levels are differentially altered in the endometrium and endometriotic lesions and to evaluate the performance of the CALD1 gene and caldesmon protein as potential biomarkers for endometriosis. Paired biopsies of endometrial tissue (eutopic endometrium) and endometriotic lesions (ectopic endometrium) were obtained from patients with endometriosis to evaluate CALD1 gene expression and caldesmon protein levels by real-time PCR and Western blot analysis, respectively. In addition, immunostaining for caldesmon to determine cellular localization was also performed. Endometrium from women without endometriosis was used as a control. Increased CALD1 expression and caldesmon levels were detected in the endometriotic lesions. The electrophoretic profile of caldesmon by Western blot analysis was clearly different between the control group (endometrium of women without endometriosis) and the group of women with endometriosis (eutopic endometrium and endometriotic lesions). Caldesmon expression as determined by immunostaining showed no variation among the cell types in endometriotic lesions and eutopic endometrium. Stromal cells marked positively in eutopic endometrium from control patients and in the endometriotic lesions. The presence of caldesmon in the endometrium of patients with and without endometriosis permitted diagnoses with 95% sensitivity (specificity 100%) and 100% sensitivity (specificity 100%) for the disease and for minimal to mild endometriosis in the proliferative phase of the menstrual cycle, respectively. In the secretory phase, minimal to mild endometriosis was detected with 90% sensitivity and 93.3% specificity. Caldesmon is a possible predictor of endometrial dysregulation in patients with endometriosis. A potential limitation of our study is the fact that other endometrial diseases were not excluded, and therefore prospective studies are needed to confirm the potential of caldesmon as a biomarker exclusively for endometriosis.
Assuntos
Proteínas de Ligação a Calmodulina/metabolismo , Endometriose/diagnóstico , Endométrio/metabolismo , Doenças Ovarianas/diagnóstico , Doenças Peritoneais/diagnóstico , Adulto , Biomarcadores/metabolismo , Proteínas de Ligação a Calmodulina/genética , Estudos de Casos e Controles , Endometriose/genética , Endometriose/metabolismo , Endométrio/patologia , Feminino , Humanos , Doenças Ovarianas/genética , Doenças Ovarianas/metabolismo , Doenças Peritoneais/genética , Doenças Peritoneais/metabolismo , Valor Preditivo dos Testes , Reação em Cadeia da Polimerase em Tempo Real , Sensibilidade e Especificidade , Células Estromais/metabolismo , Células Estromais/patologiaRESUMO
Endometriosis is a complex and multifactorial disease. Chromosomal imbalance screening in endometriotic tissue can be used to detect hot-spot regions in the search for a possible genetic marker for endometriosis. The objective of the present study was to detect chromosomal imbalances by comparative genomic hybridization (CGH) in ectopic tissue samples from ovarian endometriomas and eutopic tissue from the same patients. We evaluated 10 ovarian endometriotic tissues and 10 eutopic endometrial tissues by metaphase CGH. CGH was prepared with normal and test DNA enzymatically digested, ligated to adaptors and amplified by PCR. A second PCR was performed for DNA labeling. Equal amounts of both normal and test-labeled DNA were hybridized in human normal metaphases. The Isis FISH Imaging System V 5.0 software was used for chromosome analysis. In both eutopic and ectopic groups, 4/10 samples presented chromosomal alterations, mainly chromosomal gains. CGH identified 11q12.3-q13.1, 17p11.1-p12, 17q25.3-qter, and 19p as critical regions. Genomic imbalances in 11q, 17p, 17q, and 19p were detected in normal eutopic and/or ectopic endometrium from women with ovarian endometriosis. These regions contain genes such as POLR2G, MXRA7 and UBA52 involved in biological processes that may lead to the establishment and maintenance of endometriotic implants. This genomic imbalance may affect genes in which dysregulation impacts both eutopic and ectopic endometrium.