Relation between progesterone receptor gene polymorphism, race, parity, and uterine leiomyoma occurrence

PURPOSE: to analyze race, parity and presence of the progesterone receptor polymorphism, named PROGINS, as factors related to uterine leiomyoma occurrence in Brazilian women. METHODS: we carried out a case-control study, composed of 122 patients with the diagnosis of uterine fibroid and 125 women without the disease. After recording the clinical data, we collected biological material for DNA extraction, polymerase chain reaction and agarose gel electrophoresis in order to identify the presence of PROGINS polymorphism. Statistical analysis was performed using the non-parametric Mann-Whitney test or the chi2 test, depending on the studied variable. The risk for the occurrence of the disease was calculated by the logistic regression model, providing the odds ratio (OR). The adopted significance level was 5% (p<0.05) and the confidence interval was 95% (95% CI). RESULTS: we observed a higher prevalence of non-whitewomen - mulatto and black - (50 vs 22.4%) and nulliparas (23.8 vs 11.2%) in the cases, while the progesterone receptor genotype was more often PROGINS positive - heterozygous or mutant homozygous - among the controls (21.6 vs 10.7%). The OR indicated an elevated risk for leiomyoma related to the non-whiterace (OR=3.46; 95% CI: 2.0-6.0) and the nulliparity (OR=3.30; 95% CI: 1.9-5.6), with reduction in the presence of PROGINS-positive genotypes (OR=0.43; 95% CI: 0.2-0.9). CONCLUSIONS: the non-whiterace and nulliparity were considered risk factors for the occurrence of uterine fibroid in the studied population, while PROGINS polymorphism showed to be a protective factor.OBJETIVOS: analisamos raça, paridade e presença do polimorfismo do gene do receptor de progesterona, denominado PROGINS, como fatores relacionados à ocorrência de leiomioma uterino em mulheres brasileiras. MÉTODOS: realizamos estudo caso-controle, no qual foram incluídas 122 pacientes com diagnóstico de leiomioma e 125 mulheres sem a doença. Após registro dos dados clínicos, coletamos material biológico para extração de DNA, reação em cadeia da polimerase e eletroforese em gel de agarose, a fim de identificar a presença do polimorfismo PROGINS. A análise estatística foi feita pelo teste não paramétrico de Mann-Whitney ou pelo teste do chi2, a depender da variável estudada. O risco para ocorrência da doença foi calculado pelo modelo de regressão logística, com obtenção da odds ratio (OR) (razão de chances). O nível de significância adotado foi de 5% (p<0,05) e o intervalo de confiança foi de 95% (IC 95%). RESULTADOS: observamos maior prevalência de não-brancas- pardas e negras - (50 vs 22,4%) e de nulíparas (23,8 vs 11,2%) nos casos, ao passo que o genótipo do receptor de progesterona foi mais freqüentemente PROGINS positivo - heterozigoto ou homozigoto mutante - entre os controles (21,6 vs 10,7%). A razão de chances indicou elevação do risco para leiomioma relacionada à raça não branca(OR=3,46; IC 95%: 2,0-6,0) e à nuliparidade (OR=3,30; IC 95%: 1,9-5,6), com redução na presença de genótipos PROGINS positivo (OR=0,43; IC 95%: 0,2-0,9). CONCLUSÕES: a raça não brancae a nuliparidade foram consideradas fatores de risco para a ocorrência de leiomioma uterino em mulheres da população estudada, ao passo que o polimorfismo PROGINS demonstrou ser fator protetor.Universidade Federal de São Paulo (UNIFESP) Departamento de Ginecologia Setor de Leiomioma UterinoUniversidade Federal de São Paulo (UNIFESP) Departamento de Ginecologia Laboratório de Ginecologia MolecularUNIFESP Departamento de GinecologiaUNIFESP, Depto. de Ginecologia Setor de Leiomioma UterinoUNIFESP, Depto. de Ginecologia Laboratório de Ginecologia MolecularUNIFESP, Depto. de GinecologiaSciEL

In vivo regulation of gene expression in the corpus cavernosum by the Phoneutria nigriventer toxin

INTRODUÇÃO: O aracnídeo Phoneutria nigriventer, também conhecido por aranha-armadeira, possui um veneno complexo, contendo vários peptídeos que ativam canais iônicos nas células. Dentre estes, só dois neuropeptídeos, Tx2-5 e Tx2-6, destacam-se por relaxar o músculo liso trabecular do corpo cavernoso, induzindo ereção peniana em camundongos e ratos. Este efeito tem sido associado à produção de oxido nítrico pela ativação de óxido nítrico sintases. No entanto, faltam estudos mais amplos para determinar o papel de Tx2-6 na indução da ereção. OBJETIVOS: Identificar os genes diferencialmente expressos no tecido erétil de camundongos após indução da ereção pela Tx2-6 utilizando microarranjos de oligonucleotídeos. Validação dos resultados obtidos nos microarranjos por PCR quantitativa e imuno-histoquímica. MATERIAIS E MÉTODOS: Camundongos machos e adultos da linhagem Swiss foram divididos em dois grupos: controle (n=10), inoculados pela via intracavernosa com 20 l de solução salina; e tratado (n=10), os quais receberam 0,006gg/animal do peptídeo Tx2-6 diluído em 20 l de salina pela via intracavernosa. Uma hora após o início da ereção no grupo tratado todos os animais foram sacrificados e retirou-se o pênis. Este último foi dividido em dois fragmentos, uma parte do material foi congelada em nitrogênio líquido e mantida a 80°C até a extração do RNA para os experimentos de microarranjos e PCR quantitativa; outra parte foi utilizada para avaliação imuno-histoquímica. RESULTADOS: No grupo tratado a ereção foi observada 30-45 minutos após aplicação de Tx2-6 e mantida durante 120 minutos. Os camundongos de grupo controle não apresentaram nenhum indício de ereção. Nos experimentos de microarranjos, onde foram analisados 34.000 genes representando o genoma total do camundongo, identificou-se 3.803 (12,3%) genes com expressão diferencial de pelo menos ±1,5 vez entre os grupos (1.823 genes superexpressos e 1.980 genes subexpressos no grupo tratado comparado ao controle). Os genes ednrb, sparc, fn1, sstr2, pdgfr foram selecionados para validação dos microarranjos por PCR quantitativa e confirmaram a superexpressão em relação aos controles. As proteínas Fn1, Sstr2 e Pdgfr resultaram aumentadas no grupo tratado após avaliação imuno-histoquímica. CONCLUSÕES: A inoculação de Tx2-6 pela via intracavernosa alterou o perfil de expressão gênica no tecido erétil de camundongos. O número de genes superexpressos foi similar ao de genes subexpressos. Serão necessários outros estudos para entender melhor as vias moleculares que Tx2-6 afeta na indução da ereção peniana.INTRODUCTION: The Phoneutria nigriventer arachnid, also known as armed-spider, has a complex venom, composed by several peptides that affect cellular ionic channels. Among these, only two neuropeptides, Tx2-5 and Tx2-6 induce penile erection in mice and rats and this effect has been associated with the production of nitric oxide by the activation of nitric oxide synthases. Moreover, there is a scarcity of studies focusing on the role of Tx2-6 in the induction of erection. OBJECTIVES: To identify the differently expressed genes in the erectile tissue of mice after erection induction by Tx2-6 using oligonucleotide microarrays. To validate microarray results by quantitative PCR and immunohistochemistry. MATERIALS AND METHODS: Swiss adult male mice were divided in two groups: control (n=10) were injected intracavernously with 20 gl of saline solution; and treated (n=10) were injected intracavernously with 0.006gg/mouse of the Tx2-6 peptide diluted in 20 gl of saline solution. After checking the penile erection in the treated group, all mice were sacrificed one hour after the beginning of erection for the removal of the penis. Penile organ was divided into two fragments, one piece was immediately frozen in liquid-nitrogen and stored at -80°C until RNA extraction to make the microarray and quantitative PCR experiments; the other was reserved for immunohistochemistry analysis. RESULTS: In the treated group, erection was noticed 30-45 minutes after Tx2-6 inoculation and lasted for 120 minutes. Control mice did not present any sign of erection. Considering as differentially expressed genes with a ±1.5 fold expression difference, of the 34,000 genes on the microarray we identified 3,803 (12.3%) genes differentially expressed between the groups (1,823 genes up-regulated and 1,980 genes down-regulated in the treated group compared to controls). The ednrb, sparc, fn1, sstr2, pdgfr genes were selected for validation of microarray results by using quantitative PCR and confirmed the up-regulation when compared to controls. After immunohistochemistry analysis the Fn1, Sstr2 and Pdgfr proteins were found increased in the treated group. CONCLUSIONS: The intracavernous inoculation of Tx2-6 modified the gene expression profile of erectile tissue of mice. The number of upregulated and down-regulated genes was similar. Further studies are needed to understand the molecular pathways that Tx2-6 affect to induce penile erection

Epstein-Barr virus detection in invasive and pre-invasive lesions of the uterine cervix

In the present study, our aim was to investigate whether EBV DNA could be found in association with invasive and pre-invasive cervical cancer lesions. We hypothesize that EBV is not merely a commensal agent when present in malignant cervical lesions. DNA was extracted from cervical scrapings followed by nested PCR-based amplification. The patients were 66 women with high grade cervical intraepithelial neoplasia and 14 women with invasive cervical cancer. The control group consisted of 89 women with a normal Pap smear and colposcopy as well as a negative HPV DNA test. Analysis of our results, in conjunction with the work of other authors, leads us to propose that EBV is not merely a commensal agent when present in malignant cervical lesions. The presence of DNA from EBV is significantly associated with cervical cancer.FAPESP/Ludwig[99/03366-6

Human endometrium mRNA profile assessed by oligonucleotide three-dimensional microarray

Our purpose, in die present work, was to further comprehend the genetic events underlying the response to steroids of human endometrium from the mRNA as well as protein expression point of view. in order to achieve this goal we undertook 10 000-oligonucleotide, three-dimensional microarray analysis, followed by immunohistochemistry, on human normal endometrium in the proliferative and secretory phases of the menstrual cycle. the results revealed that a myriad of genes involved in immune response, calcium metabolism and thyroid hormone response were frequently overexpressed in the second or luteal phase of the menstrual cycle. During die follicular phase, in contrast, overexpression of genes was mainly restricted to those encoding proteins involved in cell proliferation.Universidade Federal de São Paulo, Dept Gynecol, São Paulo, BrazilUniv São Paulo, Div Urol, Lab Med Invest, São Paulo, BrazilUniv Ibirapuera, Bioodontol Post Grad Program, São Paulo, BrazilUniversidade Federal de São Paulo, Dept Gynecol, São Paulo, BrazilWeb of Scienc

Polymorphisms of the matrix metalloproteinases associated with prostate cancer

Prostate cancer (PCa) is the most common type of malignant tumor in Brazilian males. Single nucleotide polymorphisms (SNPs) have been demonstrated to be present in the promoter region of matrix metalloproteinase (MMP) genes and have been associated with the development and progression of some cancers. In this study, our aim was to investigate the association between the polymorphisms of MMP1, 2, 7, and 9 and susceptibility, and their correlation with the classic prognostic parameters of PCa. For genes MMP1, 2 and 9, the frequencies of the polymorphic homozygote genotypes were higher in the control group than in the PCa group (P<0.0001). We conclude that the MMP1, 2 and 9 polymorphisms are more common in the control group than in patients with PCa, and may have a protective effect in the development of this neoplasia

Genetic Polymorphisms of Matrix Metalloproteinases: Susceptibility and Prognostic Implications for Prostate Cancer

Purpose: Prostate cancer is the most common tumor in males in Brazil. Single nucleotide polymorphisms have been demonstrated to exist in the promoter regions of matrix metalloproteinase genes and they are associated with the development and progression of some cancers. We investigated the correlation between MMP1, 2, 7 and 9 polymorphisms with susceptibility to prostate cancer, and classic prognostic parameters of prostate cancer. Materials and Methods: Genomic DNA was extracted using conventional protocols. The DNA sequence containing the polymorphic site was amplified by realtime polymerase chain reaction using TaqMan (R) fluorescent probes. Results: For the MMP1 gene the polymorphic allele was more common in the control group than in the prostate cancer group (p <0.001). For the MMP9 gene the incidence of the polymorphic homozygote genotype was higher in the prostate cancer group (p <0.001). For higher stage tumors (pT3) a polymorphic allele in the MMP2 gene was more common (p = 0.026). When considering Gleason score, the polymorphic homozygote genotype of MMP9 was more common in Gleason 6 or less tumors (p = 0.003), while a polymorphic allele in the MMP2 gene was more common in Gleason 7 or greater tumors (p = 0.042). Conclusions: MMP1 and MMP2 may protect against prostate cancer development and MMP9 may be related to higher risk. In contrast, MMP9 polymorphism was associated with a lower Gleason score and MMP2 polymorphism was associated with nonorgan confined disease

The progesterone receptor gene polymorphism, PROGINS, may be a factor related to the development of uterine fibroids

Objective: To assess the possible association between the polymorphic allele of the progesterone receptor gene, named PROGINS, and uterine leiomyomas.Design: Case-control study.Setting: Department of Gynecology. Teaching hospital.Patient(s): One hundred twenty-two premenopausal women with fibroids and 125 postmenopausal controls not presenting the disease.Intervention(s): the subjects were classified as White or non-White (Black and Mulatto) and the progesterone receptor genotyping was performed, with DNA extracted from uterus in cases and from peripheric blood in controls and submitted to polymerase chain reaction (PCR) and agarose gel electrophoresis.Main Outcome Measure(s): the presence of the PROGINS allele was recorded, and its frequency as well as the genotypic distribution among cases and controls were compared according to race.Result(s): PROGINS-positive genotypes (heterozygous or mutant homozygous) were found in 19% of White and 11% of non-White women, and allelic frequency of PROGINS in the groups was 10.4% and 6.2%, respectively. Comparing patients and controls, we observed a significant difference among non-White women, both regarding presence of PROGINS-positive genotypes (4.9% vs. 25%, respectively), and PROGINS allele frequency (3.3% vs. 12.5%, respectively). There was no significant difference in PROGINS-positive genotypes among White cases and controls (16.4% vs. 20.6%, respectively), and in their allelic frequency (8.2% vs. 11.9%, respectively). the odds ratio showed reduced risk of fibroids related to PROGINS-positive genotypes in non-White women (odds ratio = 0.16, 95% confidence interval: 0.04-0.66), but not among White subjects (odds ratio = 0.76, 95% confidence interval: 0.33-1.74).Conclusion(s): the PROGINS polymorphism revealed to be protective in terms of uterine fibroids in Brazilian non-White women.Universidade Federal de São Paulo, Dept Gynecol, BR-04023062 São Paulo, BrazilUniversidade Federal de São Paulo, Dept Gynecol, BR-04023062 São Paulo, BrazilWeb of Scienc

Differential gene expression assessed by cDNA microarray analysis in breast cancer tissue under tamoxifen treatment

Our purpose was to identify tamoxifen (TAM) responsive genes after 30 days of TAM treatment in tumor tissues obtained from women with breast cancer using microarray expression analysis. In our study, we identified 12 candidates to be considered as tamoxifen-modulated genes. Among them, we selected two candidates the TEGT BI-1 (testis enhanced gene transcript Bax Inhibitor-1) and the CD63 gene in order to further confirm their differential expression under tamoxifen effects. We observed that both were down-regulated in tumor tissues of patients during TAM treatment. TEGT is able to inhibit the expression of Bax, which is known to promote apoptosis. On the other hand, CD63 encodes a cell membrane protein and it seems to be involved in mechanisms of platelet activation. cell adhesion and cell motility. We therefore hypothesize that TAM would be able to modulate tumor growth by down-regulating genes involved in mechanisms such as cell cycle control, tumor invasion and metastasis.Univ Fed Sao Paulo, Dept Gynecol, Mol Gynecol Lab, Paulista Sch Med, Sao Paulo, SP, BrazilLudwig Inst Canc Res, Sao Paulo Branch, Sao Paulo, BrazilUniv Fed Sao Paulo, Dept Gynecol, Mol Gynecol Lab, Paulista Sch Med, Sao Paulo, SP, BrazilWeb of Scienc