Anticancer activity of betulinic acid on MCF-7 tumors in nude mice.

Breast cancer is a major public health problem and the low effectiveness of conventional therapies to achieve long term survival results in increased mortality associated with advanced breast cancers. Betulinic acid (BA) is a pentacyclic triterpene which can be isolated from number of plants grown in the tropics. It exhibits cytotoxic activity against variety of cancer cell lines. In the present study, the in vitro cytotoxic activity and in vivo antitumor activity of BA was evaluated in athymic nude mice bearing MCF-7 breast adenocarcinoma xenografts. In vitro cytotoxic activity of BA on MCF-7 cells was studied using the MTT assay and BA was cytotoxic towards MCF-7 cells with IC50 value of 13.5µg/mL. The antitumor activity of BA was studied at concentrations of 50 and 100 mg/kg body weight in mice injected with MCF-7 cells. BA treatment delayed tumor formation and statistically significant reduction (P<0.0001) of 52 and 77% in the tumor size at concentrations of 50 and 100 mg, respectively was observed. Histopathological analysis of tumors revealed decreased angiogenesis, proliferation and invasion in BA treated animals. This is one of the first studies demonstrating the in vivo antitumor activity of BA on MCF-7 breast cancer tumors in nude mice. The antitumor effect of BA can further be enhanced by use of combination therapy and novel drug delivery systems, thus making it a promising candidate for management of breast cancer patients.

Adipocyte morphometric evaluation and angiogenesis in the omentum transposed to the breast: a preliminary study

OBJECTIVE: The purpose of this study was to describe the probable mechanism of the volume increase of laparoscopically harvested omentum flaps used to treat breast deformities. METHODS: A histological analysis of omentum samples was performed to study the volume increase of laparoscopically harvested omentum flaps. Samples were harvested immediately after the transposition of the omentum from the abdominal cavity to the breast region and during the second surgical procedure for breast symmetrization of eight patients submitted to the transposition of the omentum flap. Changes in the morphometric measurements of the adipocytes (perimeter, diameter, and area), microvascular density (as measured by the CD31 endothelial marker), and immunohistochemical expression of VEGF were documented. RESULTS: The increases in adipocyte size and microvascular density were statistically significant (P < 0.012). The expression levels of VEGF were lower in the second set of samples when compared to the first set, but the differences were not statistically significant (P < 0.093). CONCLUSION: These results demonstrate an increase in cellular volume as measured by adipocyte perimeter, diameter, and area. Moreover, the increase in the number of vessels in the second set of samples suggests that neoangiogenesis was stimulated by the initial increase in VEGF expression levels observed in the first set of samples. The increase in VEGF expression in the flap may have been caused by adipocyte hypertrophy resulting from neoangiogenesis.

D2-40, a novel lymphatic endothelial marker: identification of lymphovascular invasion and relationship with axillary metastases in breast cancer

BACKGROUND: Studies of lymphatic vessels were limited by the lack of specific markers. Recently, they have become possible due to the release of new D2-40 antibody, a selective marker for lymphatic endothelium. The aim of our study was to compare neoplastic invasion in lymphatic and blood vessels detected in hematoxylin and eosin (H&E) and immunohistochemistry-stained sections. METHODOLOGY: A total of 123 cases of invasive mammary carcinomas were studied and sorted out into three subgroups according to axillary staging (macrometastasis, micrometastasis and lymph node negative). Lymphatic vessel invasion (LVI) and blood vessel invasion (BVI) were initially evaluated in histological H&E and immunohistochemistry-stained sequential sections. Lymphatic and blood vessel invasions were assessed by immunohistochemistry, employing D2-40 and CD31 antibodies, respectively. LVI and BVI were related to size, type, histologic grade of primary tumors, and the presence of metastasis. RESULTS: LVI was detected through H&E staining procedure in 17/123 cases (13.8 percent), and through immunohistochemistry procedure in 35/123 cases (28.5 percent) (kappa = 0.433). BVI was detected through H&E in 5/123 cases (4.1 percent), and through immunohistochemistry in 19/123 cases (15.4 percent) (kappa = 0.198). LVI and BVI were positively related to higher histologic grade of primary tumors (p < 0.05). LVI was also positively related to the presence of macrometastasis. CONCLUSION: The detection of lymphatic and blood vessel invasions through immunohistochemistry employing D2-40 and CD31 was higher than the detection through H&E, and it was related to higher tumor grade and metastasis in axillary lymph nodes.

INTRODUÇÃO: Estudos de vasos linfáticos eram limitados pela ausência de marcadores endoteliais linfáticos específicos. Recentemente, eles se tornaram possíveis após liberação comercial do novo anticorpo D2-40, marcador seletivo para endotélio linfático. O objetivo do nosso estudo foi comparar invasão neoplásica em vasos linfáticos e sanguíneos detectada em secções coradas pela hematoxilina e eosina (HE) e imuno-histoquímica (IIQ). MATERIAIS E MÉTODOS: Foram estudados 123 casos de carcinomas mamários invasores subdivididos em três subgrupos de acordo com o estadiamento axilar: macrometástases (Mac-Met), micrometástases (Mic-Met) e linfonodo negativo (LNN). Invasão de vasos linfáticos (IVL) e de vasos sangüíneos (IVS) foi inicialmente avaliada em secções histológicas coradas pela HE e através da IIQ realizada em cortes seqüenciais. A invasão de vasos linfáticos e sanguíneos foi avaliada pela imuno-histoquímica, empregando-se respectivamente os anticorpos D2-40, e CD31. IVL e IVS foram relacionadas com tamanho tumoral, tipo e grau histológico dos tumores primários e com a presença de metástases. RESULTADOS: IVL foi observada pela HE em 17/123 casos (13,8 por cento) e pela IIQ em 35/123 casos (28,5 por cento) (kappa = 0,433). IVS foi observada pela HE em 5/123 casos (4,1 por cento) e pela IIQ em 19/123 casos (15,4 por cento) (kappa = 0,198). IVL e IVS estavam positivamente relacionadas com maior grau histológico dos tumores primários (p < 0,05). IVL também estava positivamente relacionada com a presença de macrometástases. CONCLUSÃO: A detecção IIQ, respectivamente por D2-40 e CD31, de invasão de vasos linfáticos e sanguíneos foi maior que a detecção feita em cortes corados pela HE e relacionou-se com maior grau tumoral e metástases em linfonodos axilares.

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.

Does color Doppler ultrasound vascularity predict the response to neoadjuvant chemotherapy in breast cancer?

OBJECTIVES: To assess the proportions of response to neoadjuvant chemotherapy of breast cancer according to color Doppler ultrasound vascularity patterns. MATERIAL AND METHOD: Prospective evaluation using gray scale and color Doppler ultrasound for number of vessels and feeding appearances of tumors as well as the changes in tumor sizes at before and after chemotherapy in 69 breast cancer patients. RESULTS: The overall response rate in 70 breast cancers was 20%. Twenty-nine lesions (41%) showed hypervascularity and 41 lesions (59%) revealed hypovascularity. There were 5 vascularity patterns and each pattern had the proportion of responders as follows; 33.3% for hypovascularity with single-vessel feeding into the tumor, 25% for hypovascularity with single-vessel feeding at periphery of the tumor, 25% for no vascular feeding to the tumor, 16.7% for hypervascularity with vascular feeding at the periphery of the tumor and 13% for hypervascularity with vascular feeding into the tumor. The highest percentage of responsive group was the pattern of hypovascularity with single-vessel feeding into the tumor (33.3%). CONCLUSION: There is a trend that tumor vascularity and patterns of vascular feeding by color Doppler ultrasound can predict the responsiveness of breast cancer to neoadjuvant chemotherapy.

Adenocarcinoma ductal infiltrante de mama en paciente masculino / Infiltrante ductual adenocarcinoma of the breast in masculine patient: a proposal of a case

Se presenta un paciente masculino, quien presentó un adenocarcinoma ductal infiltrante de mama, y quien fuera atendido en el Servicio de Cirugía 2, Servicio Autónomo Hospital Central de Maracay, Venezuela. Paciente masculino de 57 años de edad quien inició su enfermedad actual 3 meses antes de su ingreso con nódulos del 1 cm de diámetro, retroereolar izquierda, no doloroso, el cual, tiene un crecimiento rápido hasta alcanzar 5 cm x 5 cm de diámetro. Posteriormente dicha lesión se ulcera, drenando exudado serohemático en moderada cantidad, motivo por el cual consulta y le realizan biopsia que reporto carcinoma ductal infiltrante. Se realizó mastectomía radical izquierda, evidenciando lesión ulcerada de 4 cm de diámetro que infiltra hasta aponeurosis del pectoral mayor. Además de plastrón ganglionar con ganglios de 1 cm de diámetro, de consistencia aumentada. Se presenta el caso debido a lo infrecuente de encontrar adenocarcinoma de mama en un hombre. Los pacientes con cáncer de mama pueden ser sometidos a procedimientos quirúrgicos que van desde el extremo conservador de una tumorectomía hasta el opuesto de una mastectomía radical, dependiendo del criterio o preferencia del cirujano

Microvessel density in ductal carcinoma of breastand comparison with normal breast tissue

Carcinoma of the breast is the most common malignancy in adult women, and attention to it often overshadows that given other breast lesions. Angiogenesis is essential for breast cancer progression and high microvessel density [MVD] within a primary tumor has been associated with stronger likelihood of nodal metastasis and poor prognosis. In this study 50 cases of invasive ductal carcinoma of the breast were selected that had been diagnosed and archived in the Pathology Department of Mashhad Ghaem Hospital. Paraffin blocks, were cut and stained for CD34 immunohistochemical marker. Immunohistochemical slides were evaluated for microvascular density in hot spot within the tumoral and normal paratumoral tissue. Correlation between MVD with other parameters such as, age, histologic grade, lymph node involvement and tumoral sizes analyzed by parametric and nonparametric statistical evidence. MVD in normal tissue was adjusted to the tumor, and the ductal carcinoma had significant difference [12.42 +/- 2.5 and 46.76 +/- 23.4 respectivly]. MVD had relative correlation with increased histologic grade [42.3 +/- 25.1 for grade I and 63.75 +/- 36.5 for grade III]. Results of this study shows markedly differences between MVD in normal and tumoral breast tissue, also increase of histological grade have correlation with MVD increment

Tumour angiogenesis and C-myc expression in breast carcinomas.

Angiogenesis in tumors has been widely studied and has been considered to be a prognostic factor particularly in breast carcinomas. The myc family of proto oncogenes has shown an amplification in breast carcinomas. The aim of this study was to assess MVD (micro vessel density) and the expression of c-myc in invasive breast carcinomas. Formalin fixed, routinely processed, paraffin embedded tissue was stained with H&E for classification and grading according to the Nottingham modification of Bloom and Richardson criteria. Immunohistochemical staining using primary antibody against F VIII antigen and anti C-myc were carried out. LSAB method was employed with AEC as chromogen and Gill's Haematoxylin as counter stain. Micro vessels were counted and positivity indicated by a cherry red staining in the cytoplasm of the tumour cells. Grades I, II and III showed 12.22 %, 20.9% and 24.5 % positivity of micro vessel density (MVD). Fifty percent of Grade 1, 69.2 % of Grade II and 24.5 % of Grade III tumours expressed c-myc proteins. In our study we found a positive association between tumour grade and micro vessel density, signifying a poor prognosis for tumours with high MVD. C-myc expression also showed a positive association with increasing grade.

US Doppler de mama: aportes para la imagen problema / Breast Doppler US with microbuble contrast agent

Efectuamos un análisis de 20 exploraciones ecográficas de alteraciones mamarias que habían sido consideradas dudosas mediante el control clínico y mamográfico. Se utilizó la técnica del Doppler Color, pulsado y potenciado, asociando la incorporación de medio de contraste eco realzador (Levovist). La evaluación se efectuó con y sin software de medios de contraste. En todos los casos se efectuó correlación histopatológica. Ocho lesiones fueron malignas. En estas condiciones podemos decir, que el US Doppler ha demostrado habilidad para poner en evidencia otros signos de atipía, constituyéndose en un aliado de la mamografía y el US convencional

Interrelationship among prognostic factors in carcinoma breast.

Presentation of breast cancer at an advanced stage is a rule than exception in rural West Bengal. It was observed that 65.6% of fresh cases presented with history of breast lump with a duration of over 12 months. Of the total 32 cases, 27 (84.4%) had breast lump of more than 5 cm size and had 4 or more pathologically positive axillary nodes. In 28 cases (87.5%) largest node measured was 2 cm or more. In histology of breast specimen, 12 (37.5%) showed high degree of lymph vessel invasion while 31 (96.9%) had at least some lymphatic invasion. Blood vessel invasion of high degree was observed in 18 cases (56.2%) while 29 cases (90.6%) showed at least some blood vessel invasion.

Differential pattern of perivascular type IV collagen deposits in phyllodes tumors of the breast

Deposition of basement membrane extracellular matrix is influenced by adjacent tumor cells, and in some cases, the pattern of type IV collagen deposit is characteristic in malignant tumors. In this report, we analyzed the difference in type IV collagen deposition patterns between benign and malignant phyllodes tumors (PTs) of the breast. Of the 15 cases of PTs, 8 cases were benign PTs and 7 cases were malignant PTs. Three cases of other primary sarcomas of the breast (stromal sarcoma, angiosarcoma and osteosarcoma) and 2 cases of fibroadenomas were studied for comparison. The malignant PTs were distinguished from benign ones by increased mitotic figures, cellular atypism, and a higher proliferation index of stromal cells. Immunohistochemical staining against type IV collagen in malignant PTs revealed extensive to moderate deposition of type IV collagen around the small blood vessels in duplicate or multilayering pattern, while benign PTs showed minimal deposition in a single linear pattern. All of the three cases of other sarcomas revealed multilayering or meshwork pattern of type IV collagen around the blood vessels. The deposition of type IV collagen around the blood vessels may reflect the malignant behavior of the stromal tumors of the breast.