Association between matrix metalloproteinase-9 gene polymorphism and breast cancer in Brazilian women

OBJECTIVE: This study aimed to determine the relationship between rs17576 (MMP-9) polymorphism and increased cancer risk in a Brazilian breast cancer cohort. METHODS: This study included 141 women (71 breast cancer patients and 70 controls without breast cancer) who donated 3 mL of their peripheral blood for genomic DNA extraction. This DNA was then genotyped using a real-time polymerase chain reaction. RESULTS: The AG (rs17576) genotype was identified in 26 (18.43%) participants in the case group and in 22 (15.60%) participants in the control group (p=0.274), while the GG genotype was identified in ten (7.09%) participants in the case group and in one (0.70%) participant in the control group (p<0.003 - OR (95% CI) 13.13 (1.73, 593.08). No significant difference in the incidence rates was observed for AG or GG rs17576 genotypes in premenopausal women, p=0.813 and p=0.556, respectively. However, in postmenopausal women, the AG genotype was shown to occur in 14 (22.5%) participants in the case group and in 4 (6.45%) participants in the control (p<0.043), while GG genotype occurred in eight (12.90%) of the individuals in the case group and in none of the individuals in the control group (p<0.006). CONCLUSION: In this study, the MMP-9 rs17576 GG polymorphic variant was shown to be significantly associated with breast cancer risk in premenopausal women, while the AG and GG genotypes were associated with increased cancer risk in postmenopausal women.

Human Epidermal Growth Factor Receptor-2 gene polymorphism and breast cancer risk in women from the Northeastern region of Brazil

OBJECTIVES: In the Human Epidermal Growth Factor Receptor-2 (HER2) rs1136201 variant, the presence of the G allele may promote cellular alterations and increase breast cancer risk, in addition to enhanced cellular proliferation, tumor aggressiveness, and metastases. The aim of this study was to investigate the presence of the single-nucleotide polymorphism (SNP) variant, rs1136201, within the HER2 gene in women from the Northeastern region of Brazil and breast cancer risk. METHODS: The study included 140 women who were divided into two groups, case (breast cancer) and control (without breast cancer), with 70 women in each group. Peripheral blood of each woman was drawn for the study of genomic Deoxyribonucleic acid (DNA) extracted from leukocytes using the genotyping technique by real-time polymerase chain reaction. RESULTS: The GG genotype occurred in 1 woman in both groups (1.4%) (p=0.32), while the AG genotype occurred in 19 (27.2%) and 13 (18.6%) women in the case and control (p=1.00) groups, respectively. No statistically significant difference in GG and AG genotypes was observed between the case and control groups in premenopausal women (p=1.00). Furthermore, no significant difference in genotypes was observed between the groups, among postmenopausal women (p=0.14). CONCLUSION: In this study, the HER2 rs1136201 polymorphism did not show any statistically significant association with breast cancer, both in premenopausal and postmenopausal women. Nevertheless, further studies with a larger sample size should be performed to assess the association of HER2 polymorphism with breast cancer risk in women from the Northeastern region of Brazil.

Effects of anastrozole on Ki-67 antigen expression in the vaginal epithelium of female rats in persistent estrus

OBJECTIVES: Aromatase inhibitors are the first-choice drugs for the treatment of hormone sensitive breast cancer. However, in addition to the scarcity of studies, there are controversies about their effects on vaginal epithelial cell proliferation in rats, especially those in persistent estrus. METHODS: To investigate vaginal epithelial cell proliferation by Ki-67 antigen expression, persistent estrus was induced in 42 randomly selected rats. These rats were randomly divided into 2 groups: group I (control, n=21), which received 0.1 mL of propylene glycol (vehicle) daily, and group II (experimental, n=21), which received 0.5 mg/kg or 0.125 mg/day of anastrozole diluted with 0.1 mL of propylene glycol. RESULTS: Light microscopy showed a higher concentration of cells with brown Ki-67 stained nuclei in the control compared to the experimental group. The mean percentage of Ki-67 stained nuclei per 500 cells in the vaginal epithelium was 68.64±2.64 and 30.46±2.00 [mean±standard error of the mean (SEM)] in the control and experimental groups, respectively (p<0.003). CONCLUSION: This study showed that anastrozole, at the dose and treatment duration selected, significantly decreased cell proliferation in the vaginal mucosa of the rats in persistent estrus.

Evaluation of KI-67 expression in uterine leiomyoma and in healthy myometrium: a pilot study

SUMMARY OBJECTIVE Evaluate the expression of KI-67 in uterine leiomyomas and adjacent myometrial tissue and verify the existence of a correlation between clinical parameters and KI-67 expression in tumors. METHODS This is a cross-sectional, controlled, analytical study. Samples of leiomyomas and myometrium were obtained from patients who underwent hysterectomy. The samples were processed by immunohistochemistry using KI-67 antibody, and the expression was evaluated by two blinded observers. Student›s T-test was used for comparison of means, and Pearson›s P test for correlation with clinical parameters. RESULTS A total of 9 patients were included in the study. The mean age was 40.7 years, ranging from 35 to 44 years. The mean expression of KI-67 in myometrium was 1.63%, and, in leiomyomas, 5.96% (p <0.001). The highest expression of KI-67 was moderately related to the severity of anemia, bleeding, and pain level. CONCLUSION The expression of KI-67 in normal myometrium was significantly lower than in leiomyomas. The highest expression of KI-67 was moderately related to the severity of anemia, bleeding, and pain level in the patients of this study.

RESUMO OBJETIVO Avaliar a expressão do KI-67 em leiomiomas uterinos e tecido miometrial adjacente e verificar a existência de correlação entre parâmetros clínicos e expressão do KI-67 em tumores. MÉTODOS Estudo transversal, controlado e analítico. Amostras de leiomiomas e miométrio foram obtidas de pacientes que realizaram histerectomia. As amostras foram processadas por imuno-histoquímica utilizando anticorpo para KI-67 e a expressão avaliada por dois observadores cegos. O teste t de Student foi utilizado para comparação de médias e o teste P de Pearson para correlação com parâmetros clínicos. RESULTADOS Um total de 9 pacientes foi incluído no estudo. A idade média foi de 40,7 anos, variando de 35 a 44 anos. A expressão média do KI-67 no miométrio foi de 1,63% e nos leiomiomas de 5,96% (p <0,001). A maior expressão do KI-67 foi moderadamente relacionada com a gravidade da anemia, sangramento e nível de dor. CONCLUSÃO A expressão do KI-67 no miométrio normal foi significativamente menor que nos leiomiomas. A maior expressão do KI-67 foi moderadamente relacionada à gravidade da anemia, sangramento e nível de dor nos pacientes deste estudo.

Analysis of the Ki-67 index in the vaginal epithelium of castrated rats treated with tamoxifen

OBJECTIVES: Vaginal atrophy and breast cancer are common conditions in postmenopausal women and tamoxifen is the standard endocrine treatment for hormone-sensitive tumors. The present study aimed to assess the effect of tamoxifen on Ki-67 protein expression in the vaginal epithelium of castrated rats. MATERIAL AND METHODS: Forty Wistar-Hannover adult, virgin, castrated rats were randomly divided into two groups, group I (control, n=20) and group II (tamoxifen, n=20), receiving 0.5 ml of propylene glycol and 250 µg of tamoxifen diluted in 0.5 ml of propylene glycol, respectively, daily by gavage for 30 days. On the 31st day, the rats were euthanized and their vaginas were removed and fixed in 10% buffered formalin for the immunohistochemical study of Ki-67 protein expression. Data were analyzed by the Levene and Student’s t tests (p<0.05). RESULTS: The mean index of Ki-67 expression in the rat vagina of groups I and II was 4.04±0.96 and 26.86±2.19, respectively (p<0.001). CONCLUSIONS: According to the results of the present study, tamoxifen, at the dose and treatment length used, induced a significant increase in the cell proliferation of the vaginal mucosa in castrated rats, as evaluated by Ki-67 protein expression.

Aggressive angiomyxoma of the vulva: case report / Angiomixoma agressivo da vulva: relato de caso

Female patient, 42-years-old, complaining of difficulty in urinating and swelling in the vulvar area for one year. Her gynecological examination showed extensive injury in the vulvar region and the biopsy done was inconclusive. The removal of the lesion was conducted. After the procedure, the patient remains free of recurrence for 15 months. This case highlights the need to consider angiomyxoma in the differential diagnosis for tumors of unknown cause in the vulvar region.

Paciente do gênero feminino, 42 anos, com queixas de dificuldade ao urinar e aumento de volume na região vulvar há 1 ano. Ao exame ginecológico, apresentava extensa lesão na região vulvar. Biópsia da lesão foi inconclusiva. Realizou-se a exérese da lesão. A paciente permanece livre de recorrências há 15 meses. Este caso destaca a necessidade de considerar o angiomixoma no diagnóstico diferencial de massas de causa desconhecida na região vulvar.

Expressão da proteína Bax no tecido mamário normal de mulheres no menacme tratadas com raloxifeno / Expression of Bax protein in normal tissue of premenopausal women treated with raloxifene

OBJETIVO: avaliar a expressão do antígeno Bax no epitélio mamário normal de mulheres na pré-menopausa tratadas com raloxifeno. MÉTODOS: estudo randomizado duplo-cego, envolvendo 33 mulheres pré-menopáusicas com fibroadenoma. As pacientes foram divididas em dois grupos: Placebo, (n=18) e Raloxifeno 60 mg, (n=15). A medicação foi usada durante 22 dias, começando no primeiro dia do ciclo menstrual. Uma biópsia foi realizada no 23° dia do ciclo menstrual, durante a qual uma amostra do tecido mamário normal adjacente ao fibroadenoma foi coletada e submetida a estudo imuno-histoquímico utilizando o anticorpo policlonal anti-Bax para avaliar a expressão da proteína Bax. A imunorreação para a proteína Bax foi avaliada, levando-se em consideração a intensidade e a fração de células coradas, cuja combinação resultou em um escore final de 0 a 6. Os casos com escore final >3 foram classificados como positivos para proteína Bax. O teste do c2 foi usado para análise estatística dos dados (p<0,05). RESULTADOS: a porcentagem de positividade da proteína Bax foi 66,7 e 73,3 por cento nos Grupos A e B, respectivamente. Não houve diferença significante na expressão do Bax entre os dois grupos (p=0,678). CONCLUSÕES: o raloxifeno administrado por 22 dias na dose de 60 mg/dia não alterou a expressão da proteína Bax no tecido mamário normal de mulheres no menacme.

PURPOSE: to evaluate the expression of Bax antigen in the normal mammary epithelium of premenopausal women treated with raloxifene. METHODS: a randomized double-blind study was conducted in 33 ovulatory premenopausal women with fibroadenoma. Patients were divided into two groups: Placebo, (n=18) and Raloxifene 60 mg, (n=15). The medication was used for 22 days, beginning on the first day of the menstrual cycle. An excisional biopsy was carried out on the 23rd day of the menstrual cycle and a sample of normal breast tissue adjacent to the fibroadenoma was collected and submitted to immunohistochemical study using anti-Bax polyclonal antibody to evaluate the expression of Bax protein. Immunoreaction for Bax was evaluated taking into consideration intensity and fraction of stained cells, whose combination resulted in a final score ranging from 0 to 6. Cases with a final score >3 were classified as positive for Bax. The c2 test was used for statistical analysis (p<0.05). RESULTS: the percentage of positivity of Bax protein expression was 66.7 and 73.3 percent in Groups A and B, respectively. There was no significant difference in Bax expression between the two groups (p=0.678). CONCLUSIONS: raloxifene, administered for 22 days in the dose of 60 mg/day, did not alter the expression of Bax protein in the breast normal tissue of premenopausal women.

Efeitos do raloxifeno sobre a angiogênese do carcinoma de mama de mulheres menopausadas / Effects of raloxifene on angiogenesis of the breast carcinoma of menopausal women

OBJETIVO: avaliar o efeito do raloxifeno sobre a angiogênese do carcinoma de mama em mulheres menopausadas. MÉTODOS: dezesseis pacientes menopausadas com carcinoma de mama operável, estádio II (diâmetro >3 cm), positivo para receptor de estrógeno, foram incluídas no estudo. Após confirmação do diagnóstico por biópsia incisional, as pacientes receberam 60 mg de raloxifeno diariamente por 28 dias, previamente à cirurgia definitiva. Exame imunohistoquímico foi realizado nas amostras tumorais, obtidas por ocasião da biópsia para diagnóstico e avaliação do status do receptor de estrógeno e da cirurgia definitiva. O anticorpo monoclonal anti-CD34 foi usado como marcador das células endoteliais. A unidade vascular considerada foi qualquer célula ou grupo de células endoteliais coradas, nitidamente separadas de microvasos adjacentes, células tumorais ou tecido conjuntivo, formando ou não lúmen. A contagem de microvasos antes e após tratamento com raloxifeno foi realizada em dez campos aleatórios, usando microscópio acoplado a sistema de captura e análise de imagem (Imagelab®) com magnificação de 400X. O teste t de Student para duas amostras pareadas foi usado para análise estatística dos dados (p<0,05). RESULTADOS: as médias da quantidade de microvasos antes e após o tratamento com raloxifeno foi 44,4±3,5 e 22,6±1,6, respectivamente. Foi observada redução significativa da quantidade de microvasos após tratamento com raloxifeno (p<0,001). CONCLUSÕES: o tratamento primário com raloxifeno reduz significantemente a quantidade de microvasos no carcinoma de mama positivo para receptor de estrógeno em mulheres na pós-menopausa.

PURPOSE: to evaluate the effect of raloxifene on breast cancer angiogenesis of menopausal women. METHODS: sixteen menopausal women with stage II (>3 cm) estrogen receptor positive operable breast cancer were enrolled in this study. Following confirmation of the diagnosis by incisional biopsy, the patients received 60 mg raloxifene daily for 28 days prior to the definitive surgery. Immunohistochemical study was performed on the sample tumors obtained during the biopsy for the diagnosis and evaluation of the status of estrogen receptor and during the definitive surgery. The anti-CD34 monoclonal antibody was used as a marker for endothelial cells. The vascular unit was considered as any endothelial cell or group of cells of a brownish color, clearly separated from adjacent microvessels, tumor cells or other connective tissue, forming or not lumen. Microvessel count was performed in ten fields of each slide using a 40X objective lens (400X magnification). A microscope coupled to a system of capture and analysis of image was used (Imagelab®). Statistical analysis of data was carried out using the paired Student t-test and significance level was established at p<0.05. RESULTS: mean numbers of anti-CD34 antibody-stained microvessels before and after raloxifene treatment were 44.4±3.5 and 22.6±1.6, respectively. A significant reduction in the number of microvessels following raloxifene therapy was observed (p<0.001). CONCLUSIONS: when administered as primary therapy for menopausal women with breast carcinoma, raloxifene significantly reduced tumoral angiogenesis.