RESUMO
MOTIVATION: Statistical and machine-learning analyses of tumor transcriptomic profiles offer a powerful resource to gain deeper understanding of tumor subtypes and disease prognosis. Currently, prognostic gene-expression signatures do not exist for all cancer types, and most developed to date have been optimized for individual tumor types. In Galgo, we implement a bi-objective optimization approach that prioritizes gene signature cohesiveness and patient survival in parallel, which provides greater power to identify tumor transcriptomic phenotypes strongly associated with patient survival. RESULTS: To compare the predictive power of the signatures obtained by Galgo with previously studied subtyping methods, we used a meta-analytic approach testing a total of 35 large population-based transcriptomic biobanks of four different cancer types. Galgo-generated colorectal and lung adenocarcinoma signatures were stronger predictors of patient survival compared to published molecular classification schemes. One Galgo-generated breast cancer signature outperformed PAM50, AIMS, SCMGENE and IntClust subtyping predictors. In high-grade serous ovarian cancer, Galgo signatures obtained similar predictive power to a consensus classification method. In all cases, Galgo subtypes reflected enrichment of gene sets related to the hallmarks of the disease, which highlights the biological relevance of the partitions found. AVAILABILITY AND IMPLEMENTATION: The open-source R package is available on www.github.com/harpomaxx/galgo. SUPPLEMENTARY INFORMATION: Supplementary data are available at Bioinformatics online.
Assuntos
Neoplasias da Mama , Transcriptoma , Biologia Computacional , Perfilação da Expressão Gênica , Heurística , HumanosRESUMO
The importance of HSPs themselves in antigen presentation and cross-presentation remains controversial. Most studies agree that as part of their molecular chaperone function, HSPs can bind and present tumor associated antigens to professional antigen presenting cells through MHC class I and class II molecules, leading to the activation of anti-tumor CD8+ and CD4+ T cells. The regulation of the innate and adaptive immune responses by HSPs is still a matter of intense research. HSPs are seen as important anticancer vaccine adjuvants. They are used through different delivery systems: HSPs/antibodies, peptide/protein-HSP complexes, tumor antigen/HSP gene fusion, viral peptides/HSP complexes or gene fusion, viral proteins/bacterial HSP fusion. In preclinical models different administration routes, subcutaneous, intradermal, intramuscular or even peroral (under special conditions) can be used, and the animal toxicities are non-significant. The HSP-based vaccines can induce specific and non-specific cellular immune responses all of which are important to induce tumor rejection. In addition, the antibodies generated after vaccination are emerging as important protagonist in the antitumoral response. This response is significantly enhanced when the suppressive tumor microenvironment and the immune suppressing effector cells are blocked. Several clinical studies have been carried out and are ongoing, immunizing cancer patients with autologous tumor derived HSP-peptide complexes (HSPPCs). The most promising results have been observed in patients with melanoma and renal clear cell cancer without advanced disease. There are clinical trials with HSP-based anticancer vaccines other than with HSPPCs (including patients with non-Hodgkin lymphoma, high-grade transitional cell carcinoma of the bladder, high-grade cervical dysplasia, etc).
Assuntos
Vacinas Anticâncer/uso terapêutico , Proteínas de Choque Térmico/imunologia , Neoplasias/imunologia , Neoplasias/terapia , Adjuvantes Imunológicos/metabolismo , Animais , Apresentação de Antígeno , Células Apresentadoras de Antígenos/imunologia , Antígenos de Neoplasias/imunologia , Vacinas Anticâncer/imunologia , Humanos , Modelos BiológicosRESUMO
Estrogens (E) and progesterone (P) are known to require their respective steroid receptors in order to exert structural and functional effects on the oviduct. Cyclic changes in progesterone receptor (PR) localization in the oviductal tissue of female pigs were determined using an immunohistochemical technique with mouse monoclonal antibody mPRI against PR. The variations observed during the estrous cycle in the progesterone receptor (PR) intensity and proportion between ampulla and isthmus probably reflect different response of these regions to progesterone. Immediately before ovulation, during follicular phase, no staining was observed in either the ampulla or the isthmus stroma. However, a low expression of PR in the epithelium of the ampulla was observed. After ovulation, during luteal phase, PR immunostaining was more intense in the whole oviduct. According to immunohistochemical assays, the binding assays for nuclear and cytosolic PR (PRn and PRc, respectively), by using [3H] R5020 at 4 degrees C for 15 h, also showed a higher specific binding during luteal phase. However, the PR mRNA in the oviduct, analyzed by RT-PCR, showed similar levels at both stages of the estrous cycle. Although this methods could not be quantitative, indicate the possibility that a post-transcriptional control could differentially regulate the PR in the pig oviduct.
Assuntos
Tubas Uterinas/química , Fase Folicular , Expressão Gênica , Fase Luteal , Receptores de Progesterona/análise , Receptores de Progesterona/metabolismo , Suínos , Animais , Feminino , Promegestona/metabolismo , RNA Mensageiro/análise , Receptores de Progesterona/genética , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Distribuição Tecidual , TrítioRESUMO
A chronic unpredictable stress model used to produce depressive disorders in adult rats was applied to neonatal rats to investigate whether this type of stress can induce changes in the expression of Hsp70 and oestrogen receptor alpha in the oviduct, as detected by immunohistochemistry. Rats stressed during neonatal development showed changes in the expression pattern of Hsp70. In neonatal control rats, Hsp70-positive cells observed in the isthmus did not show any changes. Moreover, rats exposed to this stress model that reached adulthood had higher expression of Hsp70 in the isthmus (P<0.01) but not in the ampulla during oestrus than did the control rats. In contrast, during dioestrus, no significant changes were noted in adult rats that were stressed during neonatal development or in rats that were stressed in adulthood. These findings indicate that the isthmus is very sensitive to stressful stimuli and that repeated pre-weaning stress can change the expression of heat shock proteins in early and adult life. These subtle changes of expression in the oviduct did not affect the fertility of the rats that reached adulthood or that were mated under unstressed conditions. However, the control animals stressed during adulthood showed a disruption of the oestrous cycle: this finding is not observed in rats stressed during neonatal development that show an attenuated oestrous cycle disruption induced by chronic stress in adulthood. Moreover, there was dissociation between the expression of oestrogen receptor alpha and Hsp70. The amount of oestrogen receptor alpha remained constant in the epithelium of the oviduct in the control and in the stressed rats. Expression of oestrogen receptor alpha was noted in the stroma of the oviduct without the concomitant expression of Hsp70. It is possible that in certain cells and tissues Hsp70 is not necessary for oestrogen receptor alpha to be functional or Hsp70 might be present at very low amounts but is sufficient for the receptor to function.
Assuntos
Depressão/metabolismo , Ciclo Estral/fisiologia , Tubas Uterinas/química , Proteínas de Choque Térmico HSP70/análise , Receptores de Estrogênio/análise , Estresse Psicológico/metabolismo , Animais , Animais Recém-Nascidos , Doença Crônica , Receptor alfa de Estrogênio , Feminino , Imuno-Histoquímica/métodos , Modelos Animais , Ratos , Ratos WistarRESUMO
OBJECTIVE: To investigate Ki-67 and p21Waf1/Cip1 expression and apoptosis, before and after treatment, in tumour biopsies obtained from patients with superficial bladder cancer who underwent vinorelbine intravesical therapy. PATIENTS AND METHODS: Twenty patients with high-risk superficial bladder cancer (including one or more of the following parameters: tumour diameter > 3 cm, histological grade 3, or multicentric tumours) were treated 1-6 times (weekly) with intravesical vinorelbine (50 mg/mL) instillations. Transurethral tumour marker biopsies were obtained one week before the first instillation of the drug and one week after the last. The biopsies were immunostained for Ki-67 and p21Waf1/Cip1 with monoclonal antibodies, on tissue sections derived from paraffin-embedded samples obtained before and after vinorelbine treatments. In addition, apoptosis was determined using a terminal deoxynucleotidyl transferase-mediated dUTP biotin nick-end labelling (TUNEL) technique. RESULTS: There were no significant differences in the cell proliferation marker Ki-67 in biopsies taken before or after treatment. However, p21Waf1/Cip1 showed significantly higher expression in biopsies obtained after vinorelbine treatment, with median (range) values of 40 (20-90)% before and 70 (50-80)% after (P < 0.001, paired nonparametric Wilcoxon test). The apoptotic index was significantly higher after vinorelbine therapy, with median (range) values of 0.89 (0.06-3.8)% before and 2.25 (0.17-18.7)% after treatment (P < 0.001, paired nonparametric Wilcoxon test). Despite the brief treatment and few patients there was a clinical response in nine patients, together with low toxicity in all. CONCLUSION: The intravesical treatment of tumours with vinorelbine affects p21Waf1/Cip1 expression without blocking cell proliferation, although increasing apoptosis. The preliminary results suggest that vinorelbine may be useful for treating superficial bladder tumours, and thus a phase II study is warranted.
Assuntos
Antineoplásicos Fitogênicos/administração & dosagem , Carcinoma de Células de Transição/tratamento farmacológico , Ciclinas/metabolismo , Antígeno Ki-67/metabolismo , Proteínas de Neoplasias/metabolismo , Neoplasias da Bexiga Urinária/tratamento farmacológico , Vimblastina/administração & dosagem , Administração Intravesical , Idoso , Idoso de 80 Anos ou mais , Apoptose/efeitos dos fármacos , Biópsia/métodos , Carcinoma de Células de Transição/metabolismo , Carcinoma de Células de Transição/patologia , Inibidor de Quinase Dependente de Ciclina p21 , Feminino , Humanos , Imuno-Histoquímica/métodos , Marcação In Situ das Extremidades Cortadas , Masculino , Pessoa de Meia-Idade , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/patologia , Vimblastina/análogos & derivados , VinorelbinaRESUMO
The single-cell gel assay (comet assay) is a very useful microelectrophoretic technique for evaluation of DNA damage and repair in individual cells. Usually, the comets are visualized and evaluated with fluorescent DNA stains. This staining requires specific equipment (e.g., a high-quality fluorescence microscope), the slides must be analyzed immediately, and they cannot be stored for long periods of time. Here we describe, using human lymphocytes, some modifications of the silver staining for comets that significantly increase the sensitivity/reproducibility of the assay. This silver staining was compared with fluorescence staining and commercial silver stains. (J Histochem Cytochem 49:1183-1186, 2001)
Assuntos
Ensaio Cometa/métodos , Dano ao DNA , Linfócitos/ultraestrutura , Antineoplásicos/farmacologia , Cisplatino/farmacologia , Doxorrubicina/farmacologia , Corantes Fluorescentes , Humanos , Peróxido de Hidrogênio/farmacologia , Masculino , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Coloração pela PrataAssuntos
Humanos , Adulto , Feminino , Pessoa de Meia-Idade , Neoplasias da Mama , Resistencia a Medicamentos Antineoplásicos , Chaperonas Moleculares , Proteínas de Choque Térmico , Proteínas de Choque Térmico HSP70 , Proteínas de Choque Térmico HSP90 , Chaperonas Moleculares , Biomarcadores Tumorais , Metástase Neoplásica , Subfamília B de Transportador de Cassetes de Ligação de ATP , Proteínas de Choque Térmico , Receptores dos Hormônios Tireóideos , Proteína Supressora de Tumor p53Assuntos
Humanos , Adulto , Feminino , Pessoa de Meia-Idade , Idoso , Neoplasias da Mama/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos , Chaperonas Moleculares/efeitos dos fármacos , Proteínas de Choque Térmico/efeitos dos fármacos , Chaperonas Moleculares/diagnóstico , Proteínas de Choque Térmico/diagnóstico , Proteínas de Choque Térmico HSP70/efeitos dos fármacos , Proteínas de Choque Térmico HSP90/efeitos dos fármacos , Receptores dos Hormônios Tireóideos/efeitos dos fármacos , Proteína Supressora de Tumor p53/efeitos dos fármacos , Subfamília B de Transportador de Cassetes de Ligação de ATP/efeitos dos fármacos , Biomarcadores Tumorais , Metástase NeoplásicaRESUMO
Objetivos: En este estudio prospectivo de determinaron las modificaciones en la expresión y el valor predictivo de p53, p21 wafi/sdII/cipi, PCNA, hMLH1, hMSH2, Bcl'2 y TUNEL en pacientes con cáncer de cervix localmente avanzado tratadas con quimioterapia de inducción y radioterapia. Pacientes y métodos: Se obtuvieron muestras de 24 pacientes (IB'bulky/IIIB, 95 por ciento carcinomas escamosos) antes de la quimioterapia y a los 30 días del tratamiento. Trece pacientes recibieron un esquema de drogas basado en cisplatino y como la respuesta a esta terapia no fue buena, a las otras 11 pacientes se les administró vinorelbine e ifosfamida. Luego de la quimioterapia todas las pacientes recibieron radioterapia. La expresión de los marcadores moleculares en las biopsias pre- y post quimioterapia se estudió por inmunohistoquímica y la apoptosis fue evaluada por la técnica del TUNEL mejorada recientemente. Para comparar los cambios en la expresión de los marcadores moleculares y para correlacionarlos con la evaluación clínica (media de seguimiento: 31 meses para las pacientes que recibieron cisplatino y 19 para las que recibieron vinorelbine e ifosfamida) se realizaron análisis estadísticos. Resultados y conclusiones: La quimioterapia de inducción no aumentó la sobrevida de las pacientes, el 50 por ciento tuvo enfermedad progresiva (EP) o falleció (F). La expresión de p21waf1/sdII/cip1, hMLF1, hMSH2, y Bcl-2 no mostró cambios significativos después de la quimioterapia y no correlacionó con la evaluación clínica. La expresión de p53 no se modificó luego de la quimioterapia, las pacientes con tumores p53 positivos mostraron una tendencia a tener una sobrevida menor. Las pacientes con EP o que fallecieron mostraron niveles altos de PCNA, a diferencia de aquellas que estuvieron libres de enfermedad (LE) o con enfermedad estable (EE) (50 por ciento versus 17 por ciento, respectivamente, p<0.004). La sobrevida de las pacientes con bajos índices de TUNEL (igual o menor al valor medio entre las biopsias pre y post-quimioterapia de 1.5) fue significativamente más corta que las pacientes que presentaron índices de TUNEL altos (p<0.009). Nuestros resultados muestran que la quimioterapia de inducción (los dos tratamientos aplicados en este estudio) no mejoró la sobrevida de pacientes con cáncer de cervix...
Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Biomarcadores , Biomarcadores Tumorais , Prognóstico , Neoplasias do Colo do Útero , Apoptose , Biópsia , Genes bcl-1 , Genes bcl-2 , Imuno-Histoquímica , Marcação In Situ das Extremidades Cortadas , Biomarcadores Tumorais/isolamento & purificação , Estudos Prospectivos , Taxa de Sobrevida , Neoplasias do Colo do ÚteroRESUMO
Objetivos: En este estudio prospectivo de determinaron las modificaciones en la expresión y el valor predictivo de p53, p21 wafi/sdII/cipi, PCNA, hMLH1, hMSH2, Bcl2 y TUNEL en pacientes con cáncer de cervix localmente avanzado tratadas con quimioterapia de inducción y radioterapia. Pacientes y métodos: Se obtuvieron muestras de 24 pacientes (IBbulky/IIIB, 95 por ciento carcinomas escamosos) antes de la quimioterapia y a los 30 días del tratamiento. Trece pacientes recibieron un esquema de drogas basado en cisplatino y como la respuesta a esta terapia no fue buena, a las otras 11 pacientes se les administró vinorelbine e ifosfamida. Luego de la quimioterapia todas las pacientes recibieron radioterapia. La expresión de los marcadores moleculares en las biopsias pre- y post quimioterapia se estudió por inmunohistoquímica y la apoptosis fue evaluada por la técnica del TUNEL mejorada recientemente. Para comparar los cambios en la expresión de los marcadores moleculares y para correlacionarlos con la evaluación clínica (media de seguimiento: 31 meses para las pacientes que recibieron cisplatino y 19 para las que recibieron vinorelbine e ifosfamida) se realizaron análisis estadísticos. Resultados y conclusiones: La quimioterapia de inducción no aumentó la sobrevida de las pacientes, el 50 por ciento tuvo enfermedad progresiva (EP) o falleció (F). La expresión de p21waf1/sdII/cip1, hMLF1, hMSH2, y Bcl-2 no mostró cambios significativos después de la quimioterapia y no correlacionó con la evaluación clínica. La expresión de p53 no se modificó luego de la quimioterapia, las pacientes con tumores p53 positivos mostraron una tendencia a tener una sobrevida menor. Las pacientes con EP o que fallecieron mostraron niveles altos de PCNA, a diferencia de aquellas que estuvieron libres de enfermedad (LE) o con enfermedad estable (EE) (50 por ciento versus 17 por ciento, respectivamente, p<0.004). La sobrevida de las pacientes con bajos índices de TUNEL (igual o menor al valor medio entre las biopsias pre y post-quimioterapia de 1.5) fue significativamente más corta que las pacientes que presentaron índices de TUNEL altos (p<0.009). Nuestros resultados muestran que la quimioterapia de inducción (los dos tratamientos aplicados en este estudio) no mejoró la sobrevida de pacientes con cáncer de cervix... (AU)
Assuntos
Humanos , Feminino , Pessoa de Meia-Idade , Idoso , Neoplasias do Colo do Útero/tratamento farmacológico , Biomarcadores Tumorais/diagnóstico , Prognóstico , Biomarcadores , Estudos Prospectivos , Neoplasias do Colo do Útero/radioterapia , Biomarcadores Tumorais/isolamento & purificação , Imuno-Histoquímica , Apoptose , Taxa de Sobrevida , Biópsia , Genes bcl-1 , Genes bcl-2 , Marcação In Situ das Extremidades CortadasRESUMO
Chemosensitivity of the human colon carcinoma HCT-15 cell line to 4'-epidoxorubicin proved to be 100-fold higher than that of its variant HCT-15 EDR. Confocal scanning microscopy showed significant less drug accumulation in HCT-15 EDR. A 2-fold increase in hsp27 expression was found in HCT-15 EDR, with no alteration in hsp70. The expression of the drug exporter Pgp was similar in both cell lines, despite the lower drug accumulation shown by HCT-15 EDR in respect to HCT-15. Other molecules implicated in the acquisition of enhanced chemoresistance or a more active Pgp variant present in HCT-15 EDR, could explain the phenomenon.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Antibióticos Antineoplásicos/farmacologia , Epirubicina/farmacologia , Proteínas de Choque Térmico , Antibióticos Antineoplásicos/metabolismo , Western Blotting , Sobrevivência Celular , Neoplasias do Colo , Resistencia a Medicamentos Antineoplásicos , Ensaios de Seleção de Medicamentos Antitumorais , Epirubicina/metabolismo , Proteínas de Choque Térmico HSP27 , Proteínas de Choque Térmico HSP70/análise , Humanos , Imuno-Histoquímica , Chaperonas Moleculares , Proteínas de Neoplasias/análise , Células Tumorais CultivadasRESUMO
Certain heat shock proteins are regulated by steroid hormones and are associated with oestrogen receptor function in reproductive tissues, indicating that these proteins have a role during implantation, decidualization and placentation. In the present study, the expression of hsp25, hsp70 and oestrogen receptor alpha were examined by immunohistochemistry in oviducts from rats during neonatal development, the oestrous cycle and during early pregnancy. Oestrogen receptor alpha was the first protein observed in the neonatal oviduct, and its expression preceded that of hsp70 and hsp25. Although these heat shock proteins have been associated with the oestrogen receptor, this study showed that during early development of the oviduct, the receptor protein was not associated with the concomitant expression of hsp25 and hsp70. However, these heat shock proteins were expressed when oviductal cells became differentiated. In the adult oviduct, hsp70 was more abundant than hsp25, moreover, there were no significant modifications in expression of hsp25 during the oestrous cycle. In contrast, the expression of hsp70 was significantly higher in epithelial cells during dioestrus, when the maximum amount of oestrogen receptor alpha was also observed. Therefore, the present study shows that hsp70, but not hsp25, is an oviductal protein modulated by the oestrous cycle and that it is a protein marker for specific phases of the oestrous cycle. In addition, hsp70 was more responsive to the hormonal changes in the infundibulum and ampullar regions of the oviduct. During early pregnancy, hsp25 expression was downregulated (unlike in the endometrium), whereas hsp70 was relatively abundant in the oviduct. hsp70 was observed in all functional segments of the oviduct during pregnancy, indicating that in the oviduct, this protein is modulated by oestrogens and progesterone and possibly by other pregnancy-related hormones.
Assuntos
Animais Recém-Nascidos/crescimento & desenvolvimento , Tubas Uterinas/crescimento & desenvolvimento , Proteínas de Choque Térmico/metabolismo , Prenhez/metabolismo , Análise de Variância , Animais , Animais Recém-Nascidos/metabolismo , Western Blotting/métodos , Receptor alfa de Estrogênio , Tubas Uterinas/química , Tubas Uterinas/metabolismo , Feminino , Proteínas de Choque Térmico HSP27 , Proteínas de Choque Térmico HSP70/análise , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico/análise , Processamento de Imagem Assistida por Computador , Imuno-Histoquímica/métodos , Microscopia de Vídeo , Proteínas de Neoplasias/análise , Proteínas de Neoplasias/metabolismo , Gravidez , Ratos , Ratos Wistar , Receptores de Estrogênio/análise , Receptores de Estrogênio/metabolismoRESUMO
Tamoxifen is one of the most effective treatments for breast cancer. Standard practice is to select patients who are likely to respond to this therapy through the evaluation of estrogen receptor (ER) and progesterone receptor (PR) in the primary tumor tissue. Over the past 25 yr that physicians have been using ER determination to guide tamoxifen use, numerous studies have demonstrated that this molecular marker is useful in predicting benefit from tamoxifen. ER has been analyzed for many years using ligand-binding assays. However, current practice involves the use of immunohistochemical-based assays to detect ERalpha Immunohistochemistry (IHC) has several advantages. For example, IHC evaluates tumor cell heterogeneity, can be used to study small samples, is less expensive, and allows direct correlation with multiple histopathological tumor features and other molecular markers. PR, an estrogen-responsive protein, can also be useful in predicting response to tamoxifen in specific clinical situations. In recent years, several other markers of tamoxifen response have been examined, including: pS2 (another estrogen-regulated protein), heat-shock proteins 27 and 70, bcl-2 protein, c-erbB-2 (HER-2/neu) oncoprotein, and mutated p53 tumor suppressor protein. In this article, we present an analysis of the data on these new molecular markers. Overall, from numerous studies, the data indicate that in addition to ERalpha bcl-2 is a potential candidate to help further improve our ability to predict response to tamoxifen. ER and bcl-2 are the most useful molecular markers to better identify breast cancer patients who will respond to tamoxifen and who will have prolonged survival.
Assuntos
Biomarcadores/análise , Neoplasias da Mama/tratamento farmacológico , Tamoxifeno/uso terapêutico , Resultado do Tratamento , Neoplasias da Mama/química , Neoplasias da Mama/patologia , Proteínas de Choque Térmico/análise , Humanos , Imuno-Histoquímica , Proteínas/análise , Receptor ErbB-2/análise , Receptores de Estrogênio/análise , Receptores de Progesterona/análise , Fator Trefoil-1 , Proteína Supressora de Tumor p53/análise , Proteínas Supressoras de TumorRESUMO
We are in the process of assessing the response of cancer tissues to chemotherapy, evaluating, among other points, the proportion of cancer cells undergoing apoptosis. However, the apoptotic index obtained with the original TUNEL technique was lower than that obtained by evaluation of apoptosis on H&E-stained sections. Here we describe a small modification of the TUNEL technique that significantly increases the sensitivity of the assay. In the nonmodified TUNEL technique, a digoxigenin-labeled probe is detected using a direct peroxidase-conjugated system, whereas here we report the advantage of using a streptavidin-biotin-immunoperoxidase system. This, in conjunction with pretreatment of tissue sections with proteinase K and microwave irradiation, improved the detection of apoptotic cells.
Assuntos
Apoptose , Técnicas Imunoenzimáticas , Marcação In Situ das Extremidades Cortadas/métodos , Animais , Neoplasias da Mama/patologia , Feminino , Humanos , Masculino , Próstata/patologia , Ratos , Neoplasias do Colo do Útero/patologiaRESUMO
Expression of c-erbB-2 protein has been associated with poor prognosis and poor response to chemotherapy in breast cancer patients. In the present prospective study, we have analyzed whether c-erbB-2, p53 and P170 proteins may be determinants of tumor resistance in locally advanced breast cancer patients treated with induction chemotherapy. Biopsies (n = 60) were examined by immuno-histochemistry; in 62% of cases core or incisional biopsies were taken before drug administration, allowing comparison in paired biopsies of the cytological and molecular changes induced by treatment Sixty percent of the patients received relatively high doses of FAC or FEC (5-fluorouracil, doxorubicin or epirubicin and cyclophosphamide), and 40% received relatively high doses of doxorubicin or epirubicin alone. No significant changes were observed in the molecular markers studied following chemotherapy; in the few biopsies where changes appeared, the changes did not exhibit any significant or similar trend. For 30 of the patients who received FAC/FEC treatment, follow-up reached a median of 34 months. In these cases, neither the clinical (reduction in tumor size) nor the histological (evaluated after neoadjuvant chemotherapy) responses showed statistically significant differences between the patients who developed distant metastases and the disease-free patients. c-erbB-2 was over-expressed in 50% of patients who developed distant metastases vs. 7% of the disease-free patients. Disease free survival (DFS) curves between c-erbB-2-positive and c-erbB-2-negative patients were statistically significant. No correlation between p53 or P170 expression with DFS was found. Our results suggest that c-erbB-2 protein expression is associated with development of distant metastases in breast cancer patients treated with relatively high doses of anthracyclines in induction chemotherapy.
Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias da Mama/metabolismo , Receptor ErbB-2/metabolismo , Proteína Supressora de Tumor p53/metabolismo , Adulto , Idoso , Neoplasias da Mama/tratamento farmacológico , Ciclofosfamida/administração & dosagem , Intervalo Livre de Doença , Doxorrubicina/administração & dosagem , Resistencia a Medicamentos Antineoplásicos , Epirubicina/administração & dosagem , Feminino , Fluoruracila/administração & dosagem , Humanos , Pessoa de Meia-Idade , Estudos ProspectivosRESUMO
Heat shock proteins (Hsps) are induced in vitro by several cytotoxic drugs; in human breast cancer cells these proteins appear to be involved in anti-cancer drug resistance. The present report was designed to analyze whether chemotherapy affects in vivo the expression of Hsp27, Hsp70, Hsc70 and Hsp90 in breast cancer patients treated with induction chemotherapy and whether these proteins may be determinants of tumor resistance to drug administration. We have analyzed 35 biopsies from breast cancer patients treated with induction chemotherapy. Expression of the Hsps in the tumors was compared with (i) histological and clinical responses to chemotherapy, (ii) tumor cell proliferation measured by proliferating cell nuclear antigen (PCNA) immunostaining and nucleolar organizer regions (AgNORs) staining and (iii) the expression of estrogen and progesterone receptors. We also compared disease-free survival (DFS) and overall survival (OS) with the expression of the Hsps studied. After chemotherapy, nuclear Hsp27 and Hsp70 expression was increased and Hsp70 and Hsc70 cytoplasmic expression was decreased. A high nuclear proportion of Hsp70 in tumor cells (>10%) correlated significantly with drug resistance. We also observed that patients whose tumors expressed nuclear or a high cytoplasmic proportion (>66%) of Hsp27 had shorter DFS. The combination of Hsp27 and Hsp70 levels showed a strong correlation with DFS. Neither the cellular proliferation nor the levels of steroid receptors showed any significant difference before or after drug administration or during follow-up of patients. Our results suggest that Hsp27 and Hsp70 are involved in drug resistance in breast cancer patients treated with combination chemotherapies.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Neoplasias da Mama/química , Neoplasias da Mama/tratamento farmacológico , Resistencia a Medicamentos Antineoplásicos , Proteínas de Choque Térmico/análise , Adulto , Idoso , Protocolos de Quimioterapia Combinada Antineoplásica/administração & dosagem , Biópsia , Neoplasias da Mama/patologia , Divisão Celular , Cisplatino/administração & dosagem , Ciclofosfamida/administração & dosagem , Intervalo Livre de Doença , Doxorrubicina/administração & dosagem , Epirubicina/administração & dosagem , Fluoruracila/administração & dosagem , Proteínas de Choque Térmico HSP70/análise , Humanos , Metotrexato/administração & dosagem , Pessoa de Meia-Idade , Região Organizadora do Nucléolo/patologia , Prognóstico , Antígeno Nuclear de Célula em Proliferação/análise , Coloração pela PrataRESUMO
Heat shock protein Mr 27,000 (hsp27) is found in many human breast cancer cells and tissues; its expression is associated with the presence of estrogen receptors, lower cell proliferation, and resistance to certain chemotherapies. The purpose of this study was to assess whether hsp27 may be present in sera from women with primary breast cancer and to know whether autoantibodies to hsp27 may be found in these patients. The study was performed by Western blot analyzing sera from 42 normal premenopausal women, 20 normal postmenopausal women, and 36 breast cancer patients. hsp27 was clearly detected in sera by immunoblotting but only after immunoprecipitation. The mean hsp27 levels in cancer patients were higher than in the control patients; however, 66% of the breast cancer patients showed hsp27 within the normal range, indicating low sensitivity. Moreover, cancer patients with metastatic disease did not show significantly higher hsp27 levels than cancer patients without metastases. Serum hsp27 levels did not correlate with the hsp27 levels in tumor tissues detected by immunohistochemistry. Elevated CA 15-3 levels were not associated with high hsp27 values. Autoantibodies against hsp27 were not detected by immunoblotting in normal sera and in sera from breast cancer patients. As a consequence, serological determination of this biomarker is unlikely to be of utility in the detection and follow-up of breast cancer patients.
Assuntos
Neoplasias da Mama/sangue , Proteínas de Choque Térmico/sangue , Adulto , Análise de Variância , Biomarcadores Tumorais , Neoplasias da Mama/química , Neoplasias da Mama/imunologia , Citoplasma/metabolismo , Feminino , Proteínas de Choque Térmico/imunologia , Humanos , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Transdução de Sinais , Células Tumorais CultivadasRESUMO
In this study, we tested the hypothesis that heat shock proteins (hsps) 27 and 70 are associated with clinical resistance to tamoxifen. hsp27 is, like progesterone receptor, an estrogen-regulated protein. hsp70 is also of interest because of its interaction with estrogen receptors and because hsp70 is a component of the molecular chaperone machinery functioning in the assembly and trafficking of steroid receptors. In addition, hsps in general help protect cells against noxious stimuli and stress, and their expression has been linked to drug resistance. The study involved 205 tumors from estrogen receptor-positive tamoxifen-treated breast cancer patients with metastatic disease. All patients received daily tamoxifen as initial therapy for metastatic disease. The study began in 1982, and follow-up is now 9 years. hsp27 and hsp70 were detected by immunohistochemistry and scored according to the nuclear and/or cytoplasmic content. Expression of hsp27 or hsp70 was unrelated to estrogen receptor content, progesterone receptor content, menopausal status, age, and presence of visceral disease. Cytoplasmic and nuclear hsp27 positivities were weakly and inversely related to each other (P = 0.05). There was a significant association between cytoplasmic hsp27 and cytoplasmic hsp70 content (P < 0.001), as well as between nuclear hsp70 and nuclear hsp27 content (P = 0.001). Cytoplasmic and nuclear hsp70 were also associated (P = 0.02). However, increased hsp27 and hsp70 expression (nuclear or cytoplasmic) was not significantly associated with response to tamoxifen, time to treatment failure, or survival. Thus, this study clarifies the lack of clinical utility of hsp27 and hsp70 in predicting the response to tamoxifen in an estrogen receptor-positive breast cancer population.
Assuntos
Antineoplásicos Hormonais/uso terapêutico , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/metabolismo , Proteínas de Choque Térmico HSP70/análise , Proteínas de Choque Térmico/análise , Proteínas de Neoplasias/análise , Tamoxifeno/uso terapêutico , Adulto , Idoso , Neoplasias da Mama/patologia , Progressão da Doença , Feminino , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico/metabolismo , Humanos , Imuno-Histoquímica , Pessoa de Meia-Idade , Invasividade Neoplásica , Proteínas de Neoplasias/metabolismo , Receptores de Estrogênio/metabolismo , Estatística como Assunto , Falha de TratamentoRESUMO
In this study we have examined biopsies from women with localized primary breast cancer to investigate the prognostic performance of estrogen receptors (ER) and progesterone receptors (PR) for estimating the metastatic probability of the patients, and to explore whether discrimination gets better by combining clinicopathological and other molecular parameters into a score. This prospective study involved 205 patients with a median follow-up of 5 y. Among the evaluated clinicopathological data were: patient's age; tumor size; axillary lymph node involvement; and tumor grade. The most representative tumor samples were derived to a single laboratory for immunohistochemical evaluation of the following molecular markers: ER, PR, proliferating cell nuclear antigen (PCNA), p53 protein product, erbB-2 (HER-2/neu) oncoprotein, and P170 glycoprotein (mdrl gen product). Distant metastases (study endpoint) appeared in 19.5% (40/205) of the patients, most of these patients presented a mixture of poor, regular and good prognostic factors. Disease-free survival analysis procedures (Kaplan-Meier method) identified tumor size, axillary lymph node involvement, tumor grade, receptor status, PCNA, p53, erbB-2 and P170 as useful prognostic factors. Proportional hazard regression analysis (Cox) identified in order of importance erbB-2, tumor size, receptors status, tumor grade and PCNA as useful prognostic factors. To facilitate the evaluation of the prognostic factors, a practical and simple score system was derived. A high pathological score identified 65% of the patients that developed distant metastases, while a high molecular score was obtained in 57% of patients with metastatic disease. There was a significant improvement in the diagnosis of probability of being with distant metastases when the pathological score was combined with the molecular score, 82% of the patients with distant metastases showed an elevated combined score. Validation of this scoring system will need further larger studies (validation set as opposed to the training set used in the present study). Due to the complexity of events in cancer, the evaluation of a combination of prognostic factors should be of value to clinicians to make a more objective estimate of the prognosis of individual breast cancer patients.
Assuntos
Neoplasias da Mama/patologia , Receptores de Estrogênio/análise , Receptores de Progesterona/análise , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/análise , Análise de Variância , Protocolos de Quimioterapia Combinada Antineoplásica/uso terapêutico , Biópsia , Neoplasias da Mama/química , Neoplasias da Mama/cirurgia , Neoplasias da Mama/terapia , Quimioterapia Adjuvante , Terapia Combinada , Intervalo Livre de Doença , Feminino , Seguimentos , Humanos , Metástase Linfática , Pessoa de Meia-Idade , Análise Multivariada , Metástase Neoplásica , Probabilidade , Prognóstico , Antígeno Nuclear de Célula em Proliferação/análise , Receptor ErbB-2/análise , Taxa de Sobrevida , Tamoxifeno/uso terapêutico , Fatores de Tempo , Proteína Supressora de Tumor p53/análiseRESUMO
Human breast cancers may overexpress certain heat shock protein (hsp) family members, proteins which are involved with cell proliferation and differentiation as well as with disease prognosis and drug resistance. Here, we have studied the relationship between the expression of two hsps (hsp27 and hsp70) and the proliferative activity of tumor cells in 40 biopsies from breast cancer patients. Twenty of these tumors were selected for a detailed colocalization study. Immunocytochemistry was done using specific antibodies against hsp27 and hsp70. Cell proliferation was studied analyzing the expression of proliferating cell nuclear antigen (PCNA) (late G1, S, and G2 phases of the cell cycle) and the number of silver-staining nucleolar organizer regions (AgNORs) (G1 phase). The colocalization study revealed a statistically significant inverse correlation between hsp27 expression and cell proliferation in 16/19 (84%) of the cases evaluated by PCNA immunostaining, and in 11/16 (69%) of the cases evaluated by AgNORs. In contrast, a statistically significant positive correlation between hsp70 expression and elevated cell proliferation was seen in almost 85% of the cases evaluated by PCNA staining, and in almost 50% of the cases evaluated by AgNORs. Moreover, in 22% (9/40) of the breast cancer samples examined, hsp70 was clearly associated with the mitotic spindle. A Western blot analysis revealed that hsp70 was coprecipitated with taxol-polymerized tubulin. The association of hsp70 with the mitotic spindle was not clearly noted in lung carcinoma samples (N = 20) or in normal cells displaying elevated mitotic activity. These studies thus demonstrate that in a significant percentage of clinical breast cancers hsp27 overexpression is inversely correlated with cell proliferation, while hsp70 is clearly associated with the mitotic spindle and cell proliferation. These results add evidence to the concept that in human breast cancers hsp27 may be involved in cell growth arrest and increased differentiation while, in contrast, hsp70 may be involved in cell proliferation; further studies will be necessary to elucidate these possible cause-and-effect relationships.