The long non-coding RNA ANRASSF1 in the regulation of alternative protein-coding transcripts RASSF1A and RASSF1C in human breast cancer cells: implications to epigenetic therapy.

Alternative protein-coding transcripts of the RASSF1 gene have been associated with dual functions in human cancer: while RASSF1C isoform has oncogenic properties, RASSF1A is a tumour suppressor frequently silenced by hypermethylation. Recently, the antisense long non-coding RNA RASSF1 (ANRASSF1) was implicated in a locus-specific mechanism for the RASSF1A epigenetic repression mediated by PRC2 (Polycomb Repressive Complex 2). Here, we evaluated the methylation patterns of the promoter regions of RASSF1A and RASSF1C and the expression levels of these RASSF1 transcripts in breast cancer and breast cancer cell lines. As expected, RASSF1C remained unmethylated and RASSF1A was hypermethylated at high frequencies in 75 primary breast cancers, and also in a panel of three mammary epithelial cells (MEC) and 10 breast cancer cell lines (BCC). Although RASSF1C was expressed in all cell lines, only two of them expressed the transcript RASSF1A. ANRASSF1 expression levels were increased in six BCCs. In vitro induced demethylation with 5-Aza-2'-deoxicytydine (5-Aza-dC) resulted in up-regulation of RASSF1A and an inverse correlation with ANRASSF1 relative abundance in BCCs. However, increased levels of both transcripts were observed in two MECs (184A1 and MCF10A) after treatment with 5-Aza-dC. Overall, these findings indicate that ANRASSF1 is differentially expressed in MECs and BCCs. The lncRNA ANRASSF1 provides new perspectives as a therapeutic target for locus-specific regulation of RASSF1A.

EVALUATION OF BONE METASTASIS OF BREAST CANCER TO LONG OR SHORT BONES, ACCORDING TO MOLECULAR SUBTYPES: RETROSPECTIVE STUDY / Avaliação da metástase óssea do câncer de mama em ossos longos ou curtos, segundo os subtipos moleculares: estudo retrospectivo

Bone is the most frequent site for breast cancer metastasis. Identifying the possible preference of bone metastasis, such as long or short bones, according to molecular subtypes, could alter oncologists approach, paying special attention to these particular group of patients reducing the side effects of the bone metastatic process, involving multidisciplinary team with orthopedists, minimizing possible sequelae of this metastatic process. Detecting different metastatic sites to long or short bones, according to the molecular subtypes and their possible correlation. Fifty-eight patients with only bone metastasis were chosen. The study material was obtained from paraffin embedded primary tumors. Statistical analysis of the data was carried out. The luminal A, luminal B, hybrid luminal, HER2 + and triple-negative / basal-like molecular subtypes were identified. The molecular subtypes compared to the age of bone implants, the distribution of bone implants, and the disease free interval were not statistically significant

Acometimento ósseo é o sítio mais comum de metástase do carcinoma de mama. A identificação de possível preferência conforme os subtipos moleculares, na precocidade ou no acometimento de ossos longos ou chatos, poderia alterar a prática médica de oncologistas, dirigindo especial atenção a esses grupos de pacientes e suas possíveis complicações, em atendimento multidisciplinar com ortopedistas, minimizando possíveis sequelas desse processo metastático. Detectar a instalação dos diferentes sítios metastáticos para ossos longos ou chatos (curtos), conforme os subtipos moleculares e sua possível correlação. Foram selecionados 58 casos de pacientes com câncer de mama que apresentaram exclusivamente metástases ósseas. O material de estudo foi obtido dos tumores primários emblocados em parafina. Realizaram-se análises estatísticas dos dados. Foram identificados os subtipos moleculares luminal A, luminal B, luminal híbrido, HER2+ e triplo-negativo/basal like. Os subtipos moleculares comparados com a idade de implantes ósseos, a distribuição de implantes ósseos e o intervalo livre de doença não mostraram significância estatística.

Carcinoma intraductal bilateral em adolescente de 17 anos com ginecomastia: relato de caso / Bilateral intraductal carcinoma in a 17-year-old boy

Ginecomastia é uma alteração mamária comum na adolescência, que é, geralmente, tratadacomo problema estético. O carcinoma intraductal é uma proliferação anormal de células do revestimentodo epitélio ductal da glândula mamária, não invasiva, com potencial de desenvolverinvasão. Apresentou-se o caso de um jovem de 17 anos, que se encontrava em acompanhamentohá sete anos por ginecomastia puberal bilateral idiopática. Foi submetido à exérese de glândulasmamárias bilaterais, com resultado anatomopatológico de carcinoma intraductal dos tipos sólidoe cribiforme, de baixo grau, multifocal, com margens comprometidas, em ambas as mamas. Foiencaminhado ao Serviço de Mastologia do Hospital Amaral Carvalho, em Jaú, no estado deSão Paulo, para tratamento adicional, realizando-se mastectomia simples bilateral, sem terapiaadjuvante. Após 22 meses, encontrou-se em seguimento ambulatorial sem evidência de doençaativa. Concluiu-se que o carcinoma intraductal em homens é uma patologia rara, especialmentequando bilateral e associado à ginecomastia puberal. Por essa razão, a ginecomastia não deveser subestimada, especialmente quando a regressão espontânea não ocorrer, fazendo-se semprenecessário que o cirurgião envie a glândula mamária excisada à avaliação histológica.

Gynecomastia is a common breast change during adolescence, which is generally treated as an esthetic problem.Intraductal carcinoma is an abnormal proliferation of cells lining the ductal epithelium of the noninvasive,mammary gland, with potential to develop invasion. A case of a 17-year-old boy who was monitored for sevenyears for bilateral idiopathic pubertal gynecomastia was presented. He underwent an excision of bilateral mammaryglands with pathological results of intraductal carcinoma of solid and cribriform types, low-grade, multifocal,with positive margins in both breasts. He was referred to Serviço de Mastologia from Hospital AmaralCarvalho, in Jaú, São Paulo, for further treatment, a bilateral mastectomy was performed, without adjuvanttherapy. After 22 months, he was in follow-up without evidence of active disease. We concluded that intraductalcarcinoma in men is a rare condition, especially when bilateral and associated with pubertal gynecomastia.Therefore, gynecomastia should not be underestimated, especially when spontaneous regression does not occur,making it is always necessary for the surgeon to send the excised mammary gland to the histological evaluation.

Gene expression profile of residual breast cancer after doxorubicin and cyclophosphamide neoadjuvant chemotherapy.

In breast cancer patients, primary chemotherapy is associated with the same survival benefits as adjuvant chemotherapy. Residual tumors represent a clinical challenge, as they may be resistant to additional cycles of the same drugs. Our aim was to identify differential transcripts expressed in residual tumors, after neoadjuvant chemotherapy, that might be related with tumor resistance. Hence, 16 patients with paired tumor samples, collected before and after treatment (4 cycles doxorubicin/cyclophosphamide, AC) had their gene expression evaluated on cDNA microarray slides containing 4,608 genes. Three hundred and eighty-nine genes were differentially expressed (paired Student's t-test, pFDR<0.01) between pre- and post-chemotherapy samples and among the regulated functions were the JNK cascade and cell death. Unsupervised hierarchical clustering identified one branch comprising exclusively, eight pre-chemotherapy samples and another branch, including the former correspondent eight post-chemotherapy samples and other 16 paired pre/post-chemotherapy samples. No differences in clinical and tumor parameters could explain this clustering. Another group of 11 patients with paired samples had expression of selected genes determined by real-time RT-PCR and CTGF and DUSP1 were confirmed more expressed in post- as compared to pre-chemotherapy samples. After neoadjuvant chemotherapy some residual samples may retain their molecular signature while others present significant changes in their gene expression, probably induced by the treatment. CTGF and DUSP1 overexpression in residual samples may be a reflection of resistance to further administration of AC regimen.

Carcinoma lobular invasivo de mama com pólipo endometrial metastático / Breast invasive lobular carcinoma with metastatic endometrial polyp

Tumores extragenitais com metástase para o útero são raros, e destes o mais comum é o da mama. O tamoxifeno é uma droga utilizada como terapia adjuvante em mulheres com câncer de mama. Trata-se de droga antiestrogênica; no entanto, no endométrio sua ação é agonista para receptores de estrogênio. Esta descrição de caso relata rara associação do uso do tamoxifeno em paciente com câncer de mama que apresentou metástase para pólipo endometrial. Trata-se de paciente de 70 anos, em hormonioterapia com tamoxifeno há 19 meses, com queixa de sangramento vaginal. Em histeroscopia, evidenciaram-se pólipo endometrial e sua exérese, com diagnóstico de adenocarcinoma metastático, com provável sítio primário em mama. A paciente havia sido submetida à mastectomia radical modificada há três anos, seguida de adjuvância. Pólipos endometriais são achados comuns em mulheres menopausadas e também são complicações da terapia com tamoxifeno. A metástase de carcinoma mamário em pólipo endometrial é rara. Na literatura, foram descritas as seguintes metástases em pólipos endometriais: cinco casos por carcinoma lobular invasivo, três casos por carcinoma ductal e um caso do carcinoma apócrino. Portanto, existe dificuldade em determinar o prognóstico para essas pacientes.

Extragenital tumors metastasizing to the uterine corpus is uncommon; out of those, the most common primary site is the breast. Tamoxifen is used as adjuvant treatment for breast carcinoma. It is an antiestrogenic drug; however has a partial agonist effect on estrogen receptor in the endometrium. This case report relates rare association between the use of tamoxifen in patient with breast cancer which presented metastasis to endometrial polyps. A 10 years old woman received tamoxifen for 19 months after surgery treatment and presented vaginal bleeding. She underwent hysteroscopy with biopsy, which showed endometrial polyp. The polyp was taken out and the pathological diagnosis was metastasis from breast adenocarcinoma. Endometrial polyps are found relatively common in postmenopausal women and they are also related complications to the tamoxifen therapy. Breast carcinoma metastasis to endometrial polyp is rare. There were some descriptions in the literature: five cases of invasive lobular carcinoma, three cases of ductal carcinoma and one case of the apocrine carcinoma. There are few cases shown in the literature and, therefore, there are difficulties in determining the prognosis of these patients.

Angiossarcoma de mama após tratamento com cirugia conservadora e radioterapia / Breast angiossarcoma after conservative surgery and radiotherapy

Dois cirúrgico conservador para carcinoma ductal invasor seguido de radioterapia, vieram a desenvolver lesão violácea em mama operada e irradiada, cujo diagnóstico foi angiossarcoma, sendo uma bem diferenciada e outra pouco diferenciada. Trata-se de neoplasia rara de mama, cujo relatos de casos referentes a pacientes com média de idade de 81 anos, que, após tratamento tratamento tem base mais no procedimento cirúrgico, no qual a quimioterapia e a radioterapia não têm sua indicação bem estabelecida e clara. O prognóstico é desfavorável, com média de sobrevida de 15,5 meses.

This study features two case reports of patients with the average age of 81 years, that after conservative surgical therapy for invasive ductal carcinoma followed by radiotherapy developed purple nodule in the operated irradiated breast, whose diagnosis was angiosarcoma, being one well-differentiated and other little differentiated. It is a rare breast neoplasm, whose treatment relies more on surgery when chemotherapy and radiotherapy dont’t have well established and cleary indication. The prognostic is unfavorable, with survival prospect of 15,5 months.

Shorter CAG repeat in the AR gene is associated with atypical hyperplasia and breast carcinoma.

BACKGROUND: Previous reports into the role of [CAG]n repeat lengths in the androgen receptor (AR) gene indicate that these may play an important part in the development and progression of breast cancer, however, knowledge regarding benign breast lesions is limited. PATIENTS AND METHODS: PCR-based GeneScan analysis was used to investigate the [CAG]n repeat length at exon 1 of the AR gene in 59 benign breast lesions (27 fibroadenomas, 18 atypical hyperplasias, and 14 hyperplasias without atypia) and 54 ductal breast carcinomas. Seventy-two cancer-free women were used as a control group. In addition, [CAG]n repeats were evaluated for the presence of loss of heterozygosity (LOH) and microsatellite instability (MSI) in a subset of these samples (27 fibroadenomas, 14 hyperplasias without atypia and 22 breast carcinomas). RESULTS: Shorter [CAG]n repeat lengths were strongly correlated with atypical hyperplasias (p = 0.0209) and carcinomas (p < 0.0001). LOH was found in 1/12 and 4/20 informative cases of hyperplasias without atypia and breast carcinomas, respectively. Three patients with breast carcinoma who had previously presented atypical hyperplasia showed a reduction in the [CAG]n repeat length in their carcinomas. CONCLUSION: Short [CAG]n repeat length (< or = 20) polymorphisms are strongly associated with breast carcinomas and atypical hyperplasias. Although non-significant, a subgroup of patients with breast carcinoma and genotype SS showed an association with parameters of worse outcome.

Gene expression profile associated with response to doxorubicin-based therapy in breast cancer.

PURPOSE: This study was designed to identify genes that could predict response to doxorubicin-based primary chemotherapy in breast cancer patients. EXPERIMENTAL DESIGN: Biopsy samples were obtained before primary treatment with doxorubicin and cyclophosphamide. RNA was extracted and amplified and gene expression was analyzed using cDNA microarrays. RESULTS: Response to chemotherapy was evaluated in 51 patients, and based on Response Evaluation Criteria in Solid Tumors guidelines, 42 patients, who presented at least a partial response (> or =30% reduction in tumor dimension), were classified as responsive. Gene profile of samples, divided into training set (n = 38) and independent validation set (n = 13), were at first analyzed against a cDNA microarray platform containing 692 genes. Unsupervised clustering could not separate responders from nonresponders. A classifier was identified comprising EMILIN1, FAM14B, and PBEF, which however could not correctly classify samples included in the validation set. Our next step was to analyze gene profile in a more comprehensive cDNA microarray platform, containing 4,608 open reading frame expressed sequence tags. Seven samples of the initial training set (all responder patients) could not be analyzed. Unsupervised clustering could correctly group all the resistant samples as well as at least 85% of the sensitive samples. Additionally, a classifier, including PRSS11, MTSS1, and CLPTM1, could correctly distinguish 95.4% of the 44 samples analyzed, with only two misclassifications, one sensitive sample and one resistant tumor. The robustness of this classifier is 2.5 greater than the first one. CONCLUSION: A trio of genes might potentially distinguish doxorubicin-responsive from nonresponsive tumors, but further validation by a larger number of samples is still needed.

Marcadores prognósticos e preditivos em uma amostra aleatória de pacientes brasileiras com carcinoma invasor da mama / Prognostic and predictive markers in a random sample of Brazilian patients with invasive breast carcinoma

O câncer de mama é preponderante quanto à alta incidência cumulativa por câncer em mulheres, na maioria dos países. A avaliação dos receptores hormonais (ER-alfa e PgR) é utilizada para delinear apropriadamente as opções terapêuticas no câncer de mama. O ER-beta, assim como o ER-alfa, é um fator transcricional induzido por um ligante que pode modular a expressão de genes específicos. Em nível estrutural, os dois receptores codificam um domínio de ligação ao DNA que reconhece e se liga aos elementos de resposta ao estrógeno inseridos na região promotora de genes alvo. O receptor de progesterona é uma proteína dependente de estrógeno, ou seja, é sintetizado após o estímulo de células-alvo com o estrógeno. O gene da caderina E (CDH1) está mapeado no cromossomo 16q22.1, uma região frequentemente associada com perda de heterozigose em câncer de mama em estágios avançados. A metilação diferencial das ilhas de CpG na região promotora do gene CDH1 pode ser uma via alternativa para a perda da expressão e função da caderina-E, levando à perda da integridade tecidual, um ponto crítico da progressão tumoral. A proposta deste estudo foi investigar o perfil de expressão dos genes ER-alfa, ER-beta, e PGR pela reação em cadeia da polimerase pela transcriptase reversa (RT -PCR) e o padrão de metilação do gene CDH1 pela reação em cadeia da polimerase-metilação específica (MSP) em 80 e 79 amostras de câncer de mama, respectivamente, e correlacionar os resultados com variáveis clínicas e histopatológicas. Um subgrupo de pacientes com parâmetros clínicos e ou histopatológicos de pior prognóstico mostrou expressão gênica positiva para, pelo menos, um dos receptores hormonais (ER-alfa, ER-beta e PGR) e evolução clínica favorável num seguimento clínico superior a 24 meses (pacientes livres da doença). A co-expressão dos genes ER-alfa e ER-beta foi observada em 28 casos, 12 destes apresentaram parâmetros clínicos e ou histopatológicos de pior prognóstico...

Adenomioepitelioma de mama: relato de caso / Breast adenomyoepithelioma: case report

O adenomioepitelioma é uma lesão cuja terminologia sugere ser um tumor benigno, mas observado na literatura médica com potencialidade para o aparecimento de metástases, principalmente em gânglios axilares. Os autores apresentam um caso ocorrido após lactação. A lesão foi tratada com cirurgia local e a paciente continua em segmento contínuo.