RESUMO
INTRODUCTION: Gene amplification of CCND1 is observed in a subgroup of breast cancers with poor prognosis, whereas overexpression of the protein cyclin D1 has been linked to both worse and better clinical outcome. CCND1 amplification and protein overexpression have also been associated with resistance to treatment with tamoxifen or even to a potentially detrimental effect of tamoxifen. METHODS: To clarify these challenging and partly contrasting treatment predictive and prognostic links for cyclin D1 we analysed a large cohort of postmenopausal breast cancer patients randomised to receive either adjuvant anastrozole or tamoxifen, as part of the Arimidex, Tamoxifen, Alone or in Combination (ATAC) trial. The CCND1 amplification status and protein expression of cyclin D1 were assessed by chromogenic in situ hybridisation and immunohistochemistry, respectively, in 1,155 postmenopausal, oestrogen-receptor-positive breast cancer patients included in the TransATAC substudy. RESULTS: Amplification of CCND1 was observed in 8.7% of the tumours and was associated with increased risk of disease recurrence (hazard ratio = 1.61; 95% confidence interval, 1.08 to 2.41) after adjustment for other clinicopathological parameters. In contrast, nuclear expression of cyclin D1 protein was associated with decreased recurrence rate (hazard ratio = 0.6; 95% confidence interval, 0.39 to 0.92). The intensity of nuclear or cytoplasmic expression was not of prognostic value. There was no significant interaction between cyclin D1 status and treatment efficacy, ruling out any major detrimental effect of tamoxifen in CCND1-amplified postmenopausal breast cancer. CONCLUSIONS: In summary, CCND1 amplification and low nuclear expression of cyclin D1 predicted poor clinical outcome in postmenopausal breast cancer patients treated with either anastrozole or tamoxifen. TRIAL REGISTRATION: Current Controlled Trials ISRCTN18233230.
Assuntos
Antineoplásicos Hormonais/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Ciclina D1/genética , Recidiva Local de Neoplasia/genética , Nitrilas/uso terapêutico , Tamoxifeno/uso terapêutico , Triazóis/uso terapêutico , Anastrozol , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias da Mama/metabolismo , Neoplasias da Mama/patologia , Ciclina D1/metabolismo , Feminino , Seguimentos , Amplificação de Genes , Regulação Neoplásica da Expressão Gênica , Humanos , Antígeno Ki-67/metabolismo , Pessoa de Meia-Idade , Nitrilas/administração & dosagem , Pós-Menopausa , Valor Preditivo dos Testes , Prognóstico , Receptores de Estrogênio/metabolismo , Tamoxifeno/administração & dosagem , Resultado do Tratamento , Triazóis/administração & dosagemRESUMO
BACKGROUND: Cyclin D1 is a well-characterised cell cycle regulator with established oncogenic capabilities. Despite these properties, studies report contrasting links to tumour aggressiveness. It has previously been shown that silencing cyclin D1 increases the migratory capacity of MDA-MB-231 breast cancer cells with concomitant increase in 'inhibitor of differentiation 1' (ID1) gene expression. Id1 is known to be associated with more invasive features of cancer and with the epithelial-mesenchymal transition (EMT). Here, we sought to determine if the increase in cell motility following cyclin D1 silencing was mediated by Id1 and enhanced EMT-features. To further substantiate these findings we aimed to delineate the link between CCND1, ID1 and EMT, as well as clinical properties in primary breast cancer. METHODS: Protein and gene expression of ID1, CCND1 and EMT markers were determined in MDA-MB-231 and ZR75 cells by western blot and qPCR. Cell migration and promoter occupancy were monitored by transwell and ChIP assays, respectively. Gene expression was analysed from publicly available datasets. RESULTS: The increase in cell migration following cyclin D1 silencing in MDA-MB-231 cells was abolished by Id1 siRNA treatment and we observed cyclin D1 occupancy of the Id1 promoter region. Moreover, ID1 and SNAI2 gene expression was increased following cyclin D1 knock-down, an effect reversed with Id1 siRNA treatment. Similar migratory and SNAI2 increases were noted for the ER-positive ZR75-1 cell line, but in an Id1-independent manner. In a meta-analysis of 1107 breast cancer samples, CCND1low/ID1high tumours displayed increased expression of EMT markers and were associated with reduced recurrence free survival. Finally, a greater percentage of CCND1low/ID1high tumours were found in the EMT-like 'claudin-low' subtype of breast cancer than in other subtypes. CONCLUSIONS: These results indicate that increased migration of MDA-MB-231 cells following cyclin D1 silencing can be mediated by Id1 and is linked to an increase in EMT markers. Moreover, we have confirmed a relationship between cyclin D1, Id1 and EMT in primary breast cancer, supporting our in vitro findings that low cyclin D1 expression can be linked to aggressive features in subgroups of breast cancer.
Assuntos
Neoplasias da Mama/genética , Ciclina D1/genética , Transição Epitelial-Mesenquimal/genética , Proteína 1 Inibidora de Diferenciação/genética , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/metabolismo , Linhagem Celular Tumoral , Movimento Celular/genética , Análise por Conglomerados , Ciclina D1/metabolismo , Feminino , Perfilação da Expressão Gênica , Regulação Neoplásica da Expressão Gênica , Inativação Gênica , Humanos , Proteína 1 Inibidora de Diferenciação/metabolismo , Prognóstico , Recidiva , Risco , Fator de Crescimento Transformador beta/genéticaRESUMO
INTRODUCTION: The amplification event occurring at chromosome locus 11q13, reported in several different cancers, includes a number of potential oncogenes. We have previously reported amplification of one such oncogene, namely CCND1, to be correlated with an adverse effect of tamoxifen in premenopausal breast cancer patients. Over-expression of cyclin D1 protein, however, confers tamoxifen resistance but not a tamoxifen-induced adverse effect. Potentially, co-amplification of an additional 11q13 gene, with a resulting protein over-expression, is required to cause an agonistic effect. Moreover, during 11q13 amplification a deletion of the distal 11q region has been described. In order to assess the potential impact of the deletion we examined a selected marker for this event. METHOD: Array comparative genomic hybridization analysis was employed to identify and confirm changes in the gene expression of a number of different genes mapping to the 11q chromosomal region, associated with CCND1 amplification. The subsequent protein expression of these candidate genes was then examined in a clinical material of 500 primary breast cancers from premenopausal patients who were randomly assigned to either tamoxifen or no adjuvant treatment. The protein expression was also compared with gene expression data in a subset of 56 breast cancer samples. RESULTS: Cortactin and FADD (Fas-associated death domain) over-expression was linked to CCND1 amplification, determined by fluorescence in situ hybridization, but was not associated with a diminished effect of tamoxifen. However, deletion of distal chromosome 11q, defined as downregulation of the marker Chk1 (checkpoint kinase 1), was associated with an impaired tamoxifen response, and interestingly with low proliferative breast cancer of low grade. For Pak1 (p21-activated kinase 1) and cyclin D1 the protein expression corresponded to the gene expression data. CONCLUSIONS: The results indicate that many 11q13 associated gene products are over-expressed in conjunction with cyclin D1 but not linked to an agonistic effect of tamoxifen. Finally, the deletion of distal 11q, linked to 11q13 amplification, might be an important event affecting breast cancer outcome and tamoxifen response.
Assuntos
Antineoplásicos Hormonais/farmacologia , Neoplasias da Mama/genética , Carcinoma/genética , Cromossomos Humanos Par 11/genética , Ciclina D1/genética , Resistencia a Medicamentos Antineoplásicos/genética , Moduladores de Receptor Estrogênico/farmacologia , Proteínas de Neoplasias/genética , Pré-Menopausa , Tamoxifeno/farmacologia , Adulto , Antineoplásicos Hormonais/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Carcinoma/tratamento farmacológico , Quinase 1 do Ponto de Checagem , Hibridização Genômica Comparativa , Cortactina/biossíntese , Cortactina/genética , Ciclina D1/biossíntese , Moduladores de Receptor Estrogênico/uso terapêutico , Proteína de Domínio de Morte Associada a Fas/biossíntese , Proteína de Domínio de Morte Associada a Fas/genética , Feminino , Amplificação de Genes , Perfilação da Expressão Gênica , Humanos , Invasividade Neoplásica/genética , Proteínas de Neoplasias/biossíntese , Proteínas Quinases/biossíntese , Proteínas Quinases/genética , Ensaios Clínicos Controlados Aleatórios como Assunto/estatística & dados numéricos , Deleção de Sequência , Tamoxifeno/uso terapêutico , Quinases Ativadas por p21/biossíntese , Quinases Ativadas por p21/genéticaRESUMO
The emergence of automated image analysis algorithms has aided the enumeration, quantification, and immunohistochemical analyses of tumor cells in both whole section and tissue microarray samples. To date, the focus of such algorithms in the breast cancer setting has been on traditional markers in the common invasive ductal carcinoma subtype. Here, we aimed to optimize and validate an automated analysis of the cell cycle regulator cyclin D1 in a large collection of invasive lobular carcinoma and relate its expression to clinicopathologic data. The image analysis algorithm was trained to optimally match manual scoring of cyclin D1 protein expression in a subset of invasive lobular carcinoma tissue microarray cores. The algorithm was capable of distinguishing cyclin D1-positive cells and illustrated high correlation with traditional manual scoring (κ=0.63). It was then applied to our entire cohort of 483 patients, with subsequent statistical comparisons to clinical data. We found no correlation between cyclin D1 expression and tumor size, grade, and lymph node status. However, overexpression of the protein was associated with reduced recurrence-free survival (P=.029), as was positive nodal status (P<.001) in patients with invasive lobular carcinoma. Finally, high cyclin D1 expression was associated with increased hazard ratio in multivariate analysis (hazard ratio, 1.75; 95% confidence interval, 1.05-2.89). In conclusion, we describe an image analysis algorithm capable of reliably analyzing cyclin D1 staining in invasive lobular carcinoma and have linked overexpression of the protein to increased recurrence risk. Our findings support the use of cyclin D1 as a clinically informative biomarker for invasive lobular breast cancer.
Assuntos
Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/metabolismo , Carcinoma Lobular/metabolismo , Ciclina D1/metabolismo , Processamento de Imagem Assistida por Computador/métodos , Adulto , Idoso , Idoso de 80 Anos ou mais , Algoritmos , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/mortalidade , Carcinoma Lobular/diagnóstico , Carcinoma Lobular/mortalidade , Núcleo Celular/metabolismo , Núcleo Celular/patologia , Intervalo Livre de Doença , Feminino , Amplificação de Genes , Humanos , Mastectomia , Pessoa de Meia-Idade , Invasividade Neoplásica , Estadiamento de Neoplasias , Prognóstico , Taxa de Sobrevida , Suécia/epidemiologiaRESUMO
The G-protein coupled receptor associated protein ß-arrestin-1 is crucial for the regulation of numerous biological processes involved in cancer progression, such as intracellular signaling and cell motility. The encoding gene ARRB1 is harbored in the same chromosomal region as the CCND1 gene (11q13). Amplification of CCND1, frequently encountered in breast cancer, often involves coamplification of additional oncogenes, as well as deletion of distal 11q genes. We investigated the clinical relevance of ß-arrestin-1 in breast cancer and elucidated a potential link between ß-arrestin-1 expression and CCND1 amplification. ß-Arrestin-1 protein expression was evaluated in two breast cancer patient cohorts, comprising 179 patients (cohort I) and 500 patients randomized to either tamoxifen or no adjuvant treatment (cohort II). Additionally, migration after ß-arrestin-1 overexpression or silencing was monitored in two breast cancer cell lines. Overexpression of ß-arrestin-1 reduced the migratory propensity of both cell lines, whereas silencing increased migration. In cohort I, high expression of stromal ß-arrestin-1 was linked to reduced patient survival, whereas in cohort II both high and absent stromal expression predicted a poor clinical outcome. Patients exhibiting low or moderate levels of stromal ß-arrestin-1 did not benefit from tamoxifen, in contrast to patients exhibiting absent or high expression. Furthermore, CCND1 amplification was inversely correlated with tumor cell expression of ß-arrestin-1, indicating ARRB1 gene deletion in CCND1-amplified breast cancers.
Assuntos
Arrestinas/genética , Arrestinas/metabolismo , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/genética , Células Estromais/metabolismo , Tamoxifeno/uso terapêutico , Adulto , Idoso , Idoso de 80 Anos ou mais , Biomarcadores Tumorais/genética , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/diagnóstico , Neoplasias da Mama/patologia , Linhagem Celular Tumoral , Movimento Celular/genética , Quinase 1 do Ponto de Checagem , Ciclina D1/genética , Ciclina D1/metabolismo , Feminino , Amplificação de Genes/genética , Regulação Neoplásica da Expressão Gênica/genética , Humanos , Pessoa de Meia-Idade , Prognóstico , Proteínas Quinases/genética , Proteínas Quinases/metabolismo , Células Estromais/efeitos dos fármacos , Células Estromais/patologia , Análise de Sobrevida , Tamoxifeno/farmacologia , Resultado do Tratamento , beta-Arrestina 1 , beta-ArrestinasRESUMO
Este trabalho tem como objetivo abordar o paciente com a doença AIDS já desenvolvida e com suas seqüelas, que precisam de tratamento fisioterápico nos quadros neurológico, pneumológico e reumatológico. O tratamento foi realizado em pacientes do sexo masculino com a idade de 30 a 40 anos, homossexuais e heterossexuais, com o tempo de sobrevida superior a 2 anos. O tratamento é realizado com os devidos cuidados para nao entrar em contato com os meios de contágio