Immunohistochemistry Archives - Revista Brasileira de Ginecologia e Obstetrícia
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2007;29(6):310-316
DOI 10.1590/S0100-72032007000600006
PURPOSE: to evaluate the expression of cyclooxygenase-2 (COX-2) in ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), adjacent normal stroma, and epithelium. The correlation of expression levels with nuclear grade, histological grade, presence or absence of comedonecrosis, tumor size, and patient age was also analyzed. METHODS: forty-seven surgical samples obtained from mastectomy and quadrantectomy with simultaneous DCIS and IDC, stages I and II were included. Anti-COX-2 polyclonal antibodies were used to determine enzyme expression. Samples were classified from zero to three, in accordance with number and intensity of stained cells. RESULTS: COX-2 was positively expressed in IDC, DCIS, and normal epithelium in 86.7, 84.4, and 73.3% of the cases, respectively. Concerning nuclear grade (NG), COX-2 expression was positive in 80% of cases of NG-I; in 81.5 and 78.9% of NG II, and in 88.5 and 96.1% of NG III in DCIS and IDC, respectively. COX-2 expression occurred in 78.9% of DCIS with comedonecrosis and in 89.3% without comedonecrosis. As to histological grade (HG) of IDC, COX-2 was positive in 83.3% of HG-I; 89.9% of HG-II and 80% of HG-III. Concerning tumor diameter, COX-2 was present in 86.1% of IDC cases and in 83.3% of DCIS larger than 2 cm and in 11% of IDC and DCIS tumors ≤2 cm. The age range ≥50 years presented 90% expression for IDC and 86.7% for DCIS, and the expression was 92.5% for both IDC and DCIS in patients <50 years. CONCLUSIONS: our results demonstrated high correlation between COX-2 expression in IDC, DCIS and in the normal epithelium, which is consistent with the hypothesis that COX-2 over expression is an early event in breast carcinogenesis. There was no significant correlation between COX-2 expression in IDC and DCIS and nuclear grade, histological grade, presence of comedonecrosis, age group and tumor size.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2007;29(6):310-316
DOI 10.1590/S0100-72032007000600006
PURPOSE: to evaluate the expression of cyclooxygenase-2 (COX-2) in ductal carcinoma in situ (DCIS), invasive ductal carcinoma (IDC), adjacent normal stroma, and epithelium. The correlation of expression levels with nuclear grade, histological grade, presence or absence of comedonecrosis, tumor size, and patient age was also analyzed. METHODS: forty-seven surgical samples obtained from mastectomy and quadrantectomy with simultaneous DCIS and IDC, stages I and II were included. Anti-COX-2 polyclonal antibodies were used to determine enzyme expression. Samples were classified from zero to three, in accordance with number and intensity of stained cells. RESULTS: COX-2 was positively expressed in IDC, DCIS, and normal epithelium in 86.7, 84.4, and 73.3% of the cases, respectively. Concerning nuclear grade (NG), COX-2 expression was positive in 80% of cases of NG-I; in 81.5 and 78.9% of NG II, and in 88.5 and 96.1% of NG III in DCIS and IDC, respectively. COX-2 expression occurred in 78.9% of DCIS with comedonecrosis and in 89.3% without comedonecrosis. As to histological grade (HG) of IDC, COX-2 was positive in 83.3% of HG-I; 89.9% of HG-II and 80% of HG-III. Concerning tumor diameter, COX-2 was present in 86.1% of IDC cases and in 83.3% of DCIS larger than 2 cm and in 11% of IDC and DCIS tumors ≤2 cm. The age range ≥50 years presented 90% expression for IDC and 86.7% for DCIS, and the expression was 92.5% for both IDC and DCIS in patients <50 years. CONCLUSIONS: our results demonstrated high correlation between COX-2 expression in IDC, DCIS and in the normal epithelium, which is consistent with the hypothesis that COX-2 over expression is an early event in breast carcinogenesis. There was no significant correlation between COX-2 expression in IDC and DCIS and nuclear grade, histological grade, presence of comedonecrosis, age group and tumor size.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2000;22(7):429-433
DOI 10.1590/S0100-72032000000700005
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. Fisher's 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.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2000;22(7):429-433
DOI 10.1590/S0100-72032000000700005
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. Fisher's 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.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(3):185-191
DOI 10.1590/S0100-72032003000300007
PURPOSE: to quantitatively analyze the immunoreaction of monoclonal antibody Ki-67 in the mammary epithelium adjacent to fibroadenoma of premenopausal women treated with tamoxifen, for 50 days, at doses of 5, 10 and 20 mg/day. METHODS: we studied, prospectively, the effects of tamoxifen administered for 50 days, at doses of 5, 10 e 20 mg/day, by the immunoreaction of the Ki-67 (clone Ki-S5) monoclonal antibody on mammary epithelium adjacent to fibroadenoma in premenopausal women. We studied 58 patients in a double-blind trial who were divided into four groups: Group A (n=13; placebo), Group B (n=16; 5 mg/day tamoxifen), Group C (n=14; 10 mg/day) and Group D (n=15; 20 mg/day). All patients received the medication from the first day on of the menstrual cycle and biopsy was performed on the last day of the treatment. Cells stained and not stained by the immunoreagent were counted by optical microscopy (400X) with a digital image capturing system and image analysis. RESULTS: the average percentage of stained nuclei was calculated for all groups: Group A was 2.0 with a standard error (SE) of 0.3. In Group B it was 0.7 (SE=0.2); in Group C it was 0.4 (SE=0,2) and in Group D it was 0.1 (SE=0). Statistical analysis showed significant reductions between the groups (p<0.001), and Tukey's pairwise comparison test confirmed that there was a significant increase in the immunoreaction of the monoclonal Ki-67 antibody in groups B, C and D. CONCLUSIONS: tamoxifen, administered at doses of 5, 10 and 20 mg/day for 50 days, significantly reduced the immunoreaction of monoclonal Ki-67 in the mammary epithelium of premenopausal patients and there was no significant difference between the groups that received 5, 10 and 20 mg/day tamoxifen.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(3):185-191
DOI 10.1590/S0100-72032003000300007
PURPOSE: to quantitatively analyze the immunoreaction of monoclonal antibody Ki-67 in the mammary epithelium adjacent to fibroadenoma of premenopausal women treated with tamoxifen, for 50 days, at doses of 5, 10 and 20 mg/day. METHODS: we studied, prospectively, the effects of tamoxifen administered for 50 days, at doses of 5, 10 e 20 mg/day, by the immunoreaction of the Ki-67 (clone Ki-S5) monoclonal antibody on mammary epithelium adjacent to fibroadenoma in premenopausal women. We studied 58 patients in a double-blind trial who were divided into four groups: Group A (n=13; placebo), Group B (n=16; 5 mg/day tamoxifen), Group C (n=14; 10 mg/day) and Group D (n=15; 20 mg/day). All patients received the medication from the first day on of the menstrual cycle and biopsy was performed on the last day of the treatment. Cells stained and not stained by the immunoreagent were counted by optical microscopy (400X) with a digital image capturing system and image analysis. RESULTS: the average percentage of stained nuclei was calculated for all groups: Group A was 2.0 with a standard error (SE) of 0.3. In Group B it was 0.7 (SE=0.2); in Group C it was 0.4 (SE=0,2) and in Group D it was 0.1 (SE=0). Statistical analysis showed significant reductions between the groups (p<0.001), and Tukey's pairwise comparison test confirmed that there was a significant increase in the immunoreaction of the monoclonal Ki-67 antibody in groups B, C and D. CONCLUSIONS: tamoxifen, administered at doses of 5, 10 and 20 mg/day for 50 days, significantly reduced the immunoreaction of monoclonal Ki-67 in the mammary epithelium of premenopausal patients and there was no significant difference between the groups that received 5, 10 and 20 mg/day tamoxifen.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2001;23(5):313-319
DOI 10.1590/S0100-72032001000500007
Purpose: to compare the efficiency of anti-factor VIII and anti-CD34 antibodies as vascular makers in cervical cancer, in cervical intraepithelial neoplasia and in normal cervix. Methods: using an immunohistochemical method, factor VIII-related antigen and leukocyte antigen CD34, we performed microvascular counts in 18 squamous cell carcinomas, in 15 cervical high-grade intraepithelial neoplasia, in 15 low-grade intraepithelial lesions and in 10 normal cervices. Using light microscopy we counted microvessels per 400X field in the most active areas of neovascularization with higher microvessel density in each case. Results: the average of microvessels stained with anti-CD34 in invasive carcinoma, high-grade intraepithelial lesions, low-grade intraepithelial lesions and in the normal cervices was 154, 134, 112 and 93, respectively. When we used anti-factor VIII the average was 56, 44, 33 and 30 vessels, following the same order. High-grade intraepithelial lesions and invasive carcinomas showed greater means number of vessels than normal tissue. Conclusions: the use of anti-CD34 allowed the detection of a greater number of vessels when compared to anti-factor VIII. However, we could observe that anti-factor VIII staining was able to significantly discriminate high-grade from low-grade lesions.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2001;23(5):313-319
DOI 10.1590/S0100-72032001000500007
Purpose: to compare the efficiency of anti-factor VIII and anti-CD34 antibodies as vascular makers in cervical cancer, in cervical intraepithelial neoplasia and in normal cervix. Methods: using an immunohistochemical method, factor VIII-related antigen and leukocyte antigen CD34, we performed microvascular counts in 18 squamous cell carcinomas, in 15 cervical high-grade intraepithelial neoplasia, in 15 low-grade intraepithelial lesions and in 10 normal cervices. Using light microscopy we counted microvessels per 400X field in the most active areas of neovascularization with higher microvessel density in each case. Results: the average of microvessels stained with anti-CD34 in invasive carcinoma, high-grade intraepithelial lesions, low-grade intraepithelial lesions and in the normal cervices was 154, 134, 112 and 93, respectively. When we used anti-factor VIII the average was 56, 44, 33 and 30 vessels, following the same order. High-grade intraepithelial lesions and invasive carcinomas showed greater means number of vessels than normal tissue. Conclusions: the use of anti-CD34 allowed the detection of a greater number of vessels when compared to anti-factor VIII. However, we could observe that anti-factor VIII staining was able to significantly discriminate high-grade from low-grade lesions.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(1):23-28
DOI 10.1590/S0100-72032003000100004
PURPOSE: to determine the expression of HER-2 protein in fine-needle aspirates obtained from infiltrating breast duct carcinoma and in the corresponding surgical specimens and to correlate HER-2 expression with histological grade. Methods: forty-eight smears fixed in alcohol and previously stained by the Papanicolaou technique and histological sections of corresponding paraffin-embedded tissue blocks were submitted to immunohistochemistry by the avidin-biotin-peroxidase method using a polyclonal antibody (A0485-Dako). The reactions were carried out simultaneously with antigen recovery. The results were analyzed by a semiquantitative method according to the HercepTest score. Scores of 3+ were considered to be positive. RESULTS: positivity was observed in 39.6% of the smears and in 35.4% of the respective histological sections, with almost perfect cytohistologic concordance (kappa 0.82). The method of evaluation of histological sections was found to be reproducible regarding the smears and standardized the interpretation of the results. The correlation between HER-2 positivity and histological grade was not statistically significant but there was a tendency towards HER-2 expression in less differentiated tumors. CONCLUSION: the positivity of the immunocytochemical reactions observed in smears was comparable to that obtained in the histological sections, indicating that the material obtained by fine-needle aspiration biopsies previously stained by the Papanicolaou technique can be used for the evaluation of HER-2 expression.
Summary
Revista Brasileira de Ginecologia e Obstetrícia. 2003;25(1):23-28
DOI 10.1590/S0100-72032003000100004
PURPOSE: to determine the expression of HER-2 protein in fine-needle aspirates obtained from infiltrating breast duct carcinoma and in the corresponding surgical specimens and to correlate HER-2 expression with histological grade. Methods: forty-eight smears fixed in alcohol and previously stained by the Papanicolaou technique and histological sections of corresponding paraffin-embedded tissue blocks were submitted to immunohistochemistry by the avidin-biotin-peroxidase method using a polyclonal antibody (A0485-Dako). The reactions were carried out simultaneously with antigen recovery. The results were analyzed by a semiquantitative method according to the HercepTest score. Scores of 3+ were considered to be positive. RESULTS: positivity was observed in 39.6% of the smears and in 35.4% of the respective histological sections, with almost perfect cytohistologic concordance (kappa 0.82). The method of evaluation of histological sections was found to be reproducible regarding the smears and standardized the interpretation of the results. The correlation between HER-2 positivity and histological grade was not statistically significant but there was a tendency towards HER-2 expression in less differentiated tumors. CONCLUSION: the positivity of the immunocytochemical reactions observed in smears was comparable to that obtained in the histological sections, indicating that the material obtained by fine-needle aspiration biopsies previously stained by the Papanicolaou technique can be used for the evaluation of HER-2 expression.