M.T. Seixas

Effect of a half dose of tamoxifen on proliferative activity in normal breast tissue.

Objectives: To investigate the proliferative activity of the mammary gland epithelium and plasma levels of progesterone, estradiol, prolactin, luteinizing hormone (LH), follicle‐stimulating hormone (FSH) and sex hormone‐binding globulin (SHBG) in premenopausal women treated with 10 and 20 mg of tamoxifen (TAM) for 22 days. Patients and methods: A randomized double‐blind study was performed with 43 premenopausal women with a diagnosis of fibroadenoma of the breast. The patients were divided into three groups: A (n=15, placebo); B (n=15, TAM 10 mg/day) and C (n=13, TAM 20 mg/day). They started taking an oral dose of TAM or placebo on the very first day of the menstrual cycle. Lumpectomy was performed on the 22nd day of therapy. Normal breast tissue samples were collected during surgery, immediately immersed in 10% buffered formalin, processed for routine histology and immunohistochemistry for proliferating cell nuclear antigen (PCNA) detection. Two peripheral blood samples were collected, both on the 22nd day of the menstrual cycle, in order to evaluate the hormone levels. PCNA expressing epithelial cells were quantified by using a digital program Kontron Image System KS‐300 in 1000 cells (400×). Results: The percentage of cells expressing PCNA was significantly higher in the group receiving placebo (group A, 50.3%) when compared to groups receiving TAM 10 or 20 mg/day (group B, 24.1%; and group C, 23.2%, respectively) (P<0.001). Differences between groups B and C were not significant. Levels of progesterone, estradiol and SHBG were significantly higher in B and C groups compared to group A. Increasing concentrations of FSH (P<0.0045) and lower levels of prolactin (P<0.0055) were only found in the group receiving 20 mg/day of TAM (group C). Conclusions: A 22‐day TAM therapy, either with 10 or 20 mg/day, significantly reduced the PCNA expression and therefore the proliferative activity of the normal human breast tissue. Increasing levels of estradiol, progesterone and SHBG were associated with TAM therapy at 10 or 20 mg/day. However, a significant change of the level of FSH and prolactin was reached only with a 20‐mg/day dose.

The effect of tamoxifen on PCNA expression in fibroadenomas.

Abstract:  For almost three decades, tamoxifen has been used in the adjuvant treatment of breast cancer. It has also proven effective in the chemoprophylaxis of this disease and in the treatment of cyclic mastalgia. Since a fibroadenoma is a benign hormone‐dependent neoplasm which contains estrogen receptor (ER) levels higher than in the mammary lobule, an evaluation of the effect of this drug on the proliferative activity of both the epithelium and the stroma of fibroadenomas in premenopausal women following the administration of 10 or 20 mg/day over 22 days was proposed. Forty women with fibroadenoma were selected for a randomized double‐blind trial. They had regular menstrual cycles and had received neither hormones nor become pregnant 12 months prior to this study. Patients were divided into three groups: A (n = 14; placebo), B (n = 13; tamoxifen 10 mg/day), and C (n = 13; tamoxifen 20 mg/day). The treatment was initiated on the first day of their menstrual cycle and the surgeries were performed on the 22nd day. Estradiol, progesterone, and steroid hormone binding globulin (SHBG) were measured twice. The first measurement was performed on the 22nd day of the previous menstrual cycle and the second one was performed on the day of surgery. The fibroadenoma was fixed in 10% formaldehyde solution and stained with hematoxylin and eosin and then processed through immunohistochemical reaction (PC‐10, DAKO code number M879, Denmark A/S). The immunoexpression of the proliferative cell nuclear antigen (PCNA) of at least 500 epithelial and 500 stromal cells was evaluated. Such cells were interactively counted using the Kontron Imaging System KS‐300 computerized analysis system, with ×400 magnification. As to PCNA expression in the fibroadenomas’ epithelium, the average percentage of stained nuclei in groups A, B, and C was 25.2, 19.3, and 18.0, respectively. However, no significant difference was found in the variance analysis of these data (p = 0.168). As to the study of the fibroadenomas’ stroma, the average percentage of stained nuclei found in groups A, B, and C was 32.4, 23.2, and 18.4, respectively. The variance analysis (p = 0.031) and Fishers multiple comparison test (1.39; 26.67 confidence interval [CI]) confirmed that the number of PCNA‐expressing nuclei in the stroma was significantly lower in group C (20 mg/day) compared to group A (control). However, there was no significant difference between group B (10 mg/day) and group C (20 mg/day). It was found that tamoxifen reduced the expression of PCNA in the epithelium and the stroma of the fibroadenoma. However, the effect was only statistically significant in the stroma when a 20 mg/day dose was administered.

Evaluation of Monoclonal Antibody MIB-1 in the Mammary Epithelium Adjacent to Fibroadenomas in Premenopausal Women Treated with Tamoxifen

The purpose of this study was to study the monoclonal antibody MIB‐1 in the normal breast epithelium adjacent to a fibroadenoma in women in the luteal phase of the menstrual cycle who were treated with tamoxifen at doses of 10 and 20 mg for 22 days. The proliferative activity of the mammary epithelium adjacent to the fibroadenoma was studied by immunohistochemistry on the basis of the monoclonal antibody MIB‐1 (Immunotech, catalog No. 0505, lot 001). The study was randomized and double blind and was conducted on 44 women with fibroadenomas divided into three groups: A (n = 16, placebo), B (n = 15, tamoxifen, 10 mg), and C (n = 13, tamoxifen, 20 mg). Tamoxifen was administered for 22 days starting on the 2nd day of the menstrual cycle, and a biopsy was taken on the 23rd day. Serum estradiol, progesterone, sex hormone binding globulin, follicle‐stimulating hormone, luteinizing hormone, and prolactin were measured before treatment (21st and 24th day of the previous menstrual cycle) and on the day of the biopsy. The mean percentage of stained nuclei per 1,000 cells was 9.2 in group A, 4.5 in group B, and 3.2 in group C. The Fishers test revealed that tamoxifen significantly reduced MIB‐1 at doses of 10 and 20 mg compared with the placebo group (p < 0.0001), with no significant differences between doses in terms of proliferative activity (p = 0.21). Groups B and C presented a significant increase in progesterone (p = 0.038), estradiol (p < 0.001), and sex hormone binding globulin (p = 0.001) levels. Elevation of serum follicle‐stimulating hormone concentration (p = 0.0045) and a fall in prolactin levels (p = 0.0055) were observed. We conclude that tamoxifen significantly reduced the proliferative activity of the mammary epithelium at the doses of 10 and 20 mg/day.

Sarcoma granulocítico em órbita: relato de caso

Orbital granulocytic sarcoma is a localized tumor consisting of malignant cells of myeloid origin. This tumor may present in association with acute myelogenous leukemia. Granulocytic sarcoma may be found in a variety of locations throughout the body including the orbit and typically affects children and young adults. There is a slight male predominance in these cases. This is an uncommon case report of a 33-year-old Latin-American woman who was admitted to the Hospital for rapidly progressive orbital proptosis. There was no systemic manifestation of leukemia. The occurrence of orbital granulocytic sarcoma before the development of systemic leukemia in children and young adults is not uncommon and these cases frequently develop hematological evidence within 2 months after initial orbital disease. In this case report, there was no systemic manifestation of leukemia in the last 30 months, even in the presence of orbital tumors. Granulocytic sarcoma is most frequently confused with malignant lymphoma, rhabdomyosarcoma and neuroblastoma. The differential diagnosis of these cases can be challenging, particularly when there is no evidence of systemic leukemia, when imaging features are not sufficiently specific to distinguish granulocytic neoplasms from other tumors. To establish the diagnosis often a biopsy is required. The treatment in such cases (orbital granulocytic sarcoma) is not standardized. Orbital granulocytic sarcoma may be suspected in cases of orbital tumors even in the absence of systemic manifestations of leukemia at any age.

The effect of tamoxifen on the proliferative activity of fibroadenomas

Ž . For almost three decades, tamoxifen TAM has been used in the adjuvant treatment of breast cancer 1 . It has also proven effective in the chemoprophylaxis of this disease and in the treatment of cyclical mastalgia 2,3 . As a fibroadenoma is a benign hormone-dependant neoplasm, which bears levels of estrogen receptors higher than in the mammary lobule 4 , an evaluation of the effect of this drug on the proliferative activity of both the epithelium and the stroma of fibroadenomas in premenopausal women following the administration of 10 and 20 mg day during 22 days was proposed. Forty females with fibroadenoma were selected for a randomized double-blind trial. They had regular menstrual cycles and had neither received hormones nor become pregnant 12 months prior

Avaliação da Atividade Proliferativa no Epitélio Mamário Adjacente a Fibroadenoma em Mulheres Tratadas com Tamoxifeno

Purpose: to study the monoclonal antibody MIB-1 in the normal breast epithelium adjacent to a fibroadenoma in women in the luteal phase of the menstrual cycle treated with tamoxifen. Patients and methods: the proliferative activity of the mammary epithelium adjacent to the fibroadenoma was studied by immunohistochemistry based on immunoexpression of the monoclonal antibody MIB-1. The study was randomized and double blind and was conducted on 44 women with fibroadenomas, divided into 3 groups: A (n = 16; placebo), B (n = 15; tamoxifen, 10 mg), and C (n = 13; tamoxifen, 20 mg). Tamoxifen was administered for 22 days starting on the 2nd day of the menstrual cycle and a biopsy was taken on the 23rd day. Results: the mean percentage of stained nuclei per 1000 cells was 9.2 in group A, 4.5 in group B, and 3.2 in group C. Fishers test revealed that tamoxifen significantly reduced the immunoexpression of MIB-1 at the doses of 10 and 20 mg compared to the placebo group (p<0.0001), with no significant differences between doses in terms of proliferative activity (p = 0.21). Conclusion: we conclude that tamoxifen significantly reduced the proliferative activity of the mammary epithelium at the doses of 10 and 20 mg/day.

Evaluation of monoclonal antibody MIB-1 in the mammary epithelium adjacent to fibroadenomas in premenopausal women treated with tamoxifen

The purpose of this study was to study the monoclonal antibody MIB-1 in the normal breast epithelium adjacent to a fibroadenoma in women in the luteal phase of the menstrual cycle who were treated with tamoxifen at doses of 10 and 20 mg for 22 days. The proliferative activity of the mammary epithelium adjacent to the fibroadenoma was studied by immunohistochemistry on the basis of the monoclonal antibody MIB-1 (Immunotech, catalog No. 0505, lot 001). The study was randomized and double blind and was conducted on 44 women with fibroadenomas divided into three groups: A (n=16, placebo), B (n=15, tamoxifen, 10 mg), and C (n=13, tamoxifen, 20 mg). Tamoxifen was administered for 22 days starting on the 2nd day of the menstrual cycle, and a biopsy was taken on the 23rd day. Serum estradiol, progesterone, sex hormone binding globulin, follicle-stimulating hormone, luteinizing hormone, and prolactin were measured before treatment (21st and 24th day of the previous menstrual cycle) and on the day of the biopsy. The mean percentage of stained nuclei per 1,000 cells was 9.2 in group A, 4.5 in group B, and 3.2 in group C. The Fishers test revealed that tamoxifen significantly reduced MIB-1 at doses of 10 and 20 mg compared with the placebo group (p < 0.0001), with no significant differences between doses in terms of proliferative activity (p=0.21). Groups B and C presented a significant increase in progesterone (p=0.038), estradiol (p < 0.001), and sex hormone binding globulin (p=0.001) levels. Elevation of serum follicle-stimulating hormone concentration (p=0.0045) and a fall in prolactin levels (p=0.0055) were observed. We conclude that tamoxifen significantly reduced the proliferative activity of the mammary epithelium at the doses of 10 and 20 mg/day.

Effect of taximofen on the expression of proliferative cell nuclear antigen (PCNA) in the epithlium and stroma of fibroadenomas in women during menacme

Objectives: The aim of the study was to investigate the effect of Tamoxifen on the proliferative activity of both the epithelium and the stroma of fibroadenomas in women during menacme following the administration of 10 and 20 mgiday during 22 days. Study Methods: 41 females with fibroadenoma were selected for a randomized double-blind trial. They presented regular menstrual cycles and had neither received hormones nor become pregnant 12 months prior to this study. Patients were divided in three groups: A (n=15; placebo), B (n=13; 10 mgiday) and C (n=13; 20 mgiday). The treatment was initiated on the 1”’ day of their menstrual cycle, and the surgeries were performed on the 22”d day. Estradiol, progesterone and steroid hormone binding globulin (SHBG) were measured twice. The first measurement was performed on the 22”d day of the previous menstrual cycle and the second one on the occasion of the surgery. The mammary tissue was fixed in 10% formaldehyde solution and stained with HE and then processed through immunohistochemical reaction (PC-10 DAK0 code number M879 Denmark AiS). The immunoexpression of the proliferative cell nuclear antigen (PCNA) of at least 500 epithelial and 500 stromal cells was evaluated. Such cells were interactively counted using Kontron Imaging System KS-300 computerized analysis system, with 400X magnification. Results: As to the PCNA expression in the epithelium of fibroadenomas, the average percentage of stained nuclei in groups A, B and C was 25.2 (standard error = 3.0), 19.3 (standard error = 2.9) and 18.0 (standard error = 2.5), respectively. However, no significant difference was found in the variance analysis of these data (p=O.168). Upon the study of the stroma of fibroadenomas, the average percentage of stained nuclei found in groups A, B and C was 32.4 (standard error = 3.3), 23.2 (standard error = 4.6) and 18.4 (standard error = 3.0), respectively. The variance analysis (p=O.O31) and Fisher’s multiple comparison test (1.39; 26.67 confidence interval) confirmed that the number of PCNA expressing nuclei in the stroma was significantly lower in group C (20 mgiday) when compared to group A (control). However, there was no significant difference between groups (10 mgiday) and C (20 mgiday). Conclusion: Thus, it was concluded that the administration of 10 mgiday and 20 mgiday of tamoxifen reduced the PCNA expression in the stroma of fibroadenoma after 22 days of treatment; however, its effect was statistically significant only in the stroma when doses of 20 mgiday were administered.

A study of apoptosis in normal breast tissue in tamoxifen‐treated premenopausal women with fibroadenoma

Objectives: Tamoxifen (TAMi20mgiday) has been used in breast cancer chemoprevention and exerts a long-term suppressive effect on human breast cancer cell proliferation. The aim of the study is to determine the apoptosis rate in the normal human breast epithelium adjacent to fibroadenoma during TAM treatment (10 and 20 mgiday). Study Methods: We evaluated a group of 40 premenopause fibroadenoma patients during 22 days of therapy. By using a doubleblind randomized study patients were divided according to the following groups: Group I 14 patients used as a control group. Group II 13 patients receiving 10 mg of TAM/day and Group III 13 patients receiving 20 mg TAM/day. Treatment was started on the first day of menstrual cycle and biopsies were performed on the 22”d day of therapy. Serum levels of estradiol, progesterone, prolactin, FSH, LH and SHBG were measured on the 22”d of the last cycle as well as on the biopsy day. Mammary samples were immediately fixed on 10% buffered formalin and included in paraffin. H.E. staining was performed in order to quantitate the number of apoptotic cells in 10 different fields at 400x magnification. Results: The mean number of apoptotic cells in group I was 34.8 cells. Group II (10 mgiday) 12.9 and group III (20 mgiday) 12.0 apoptotic cells. Statistical analysis showed significant reductions in groups II and III when compared to the control group. On the other hand, there were no differences between groups II and III (Fisher’s test p<O.OOl). Conclusions: According to our findings, tamoxifen (10 or 20 mgiday) reduced the apoptosis rate on the human normal breast epithelium after 22 days of treatment.