Comparison Of Expression Site Of HER2, VEGF, Ki67 And TOPO?? In Benign And Malignant Breast Diseases

Objective: Breast hyperplasia(sclerosing adenosis) and breast fibroadenoma can exist in the form of breast lumps, and their clinical manifestation were similar to breast cancer. The breast hyperplasia had close relationship to breast cancer, and extremely easy to transforming to cancer, but the malignant rate of breast fibroadenoma is very low. Why dose the same hyperplasia produce different outcomes? In this study, we selected four indexes, which were HER2 closely related to the occurrence and development of breast cancer, Ki67 ? TOPO?? related to cell proliferation, and VEGF related to cell infiltration and growth to compare the expression site in normal breast tissue, breast hyperplasia(sclerosing adenosis), fibroadenoma and breast invasive ductal carcinoma, to explore their role and significance in the development of breast cancer and to preliminarily explore the pathogenesis of breast cancer.Methods: Breast tissue samples were obtained from patients who had an operation in thoracic surgery of Baoding Second Central Hospital between June 2013 to April 2015, of which 3 cases were normal breast, 3 cases were sclerosing adenosis, 3 cases were breast fibroadenoma, 60 cases were breast cancer(Invasive ductal carcinoma of breast confirmed by surgery and pathology).Pathologic grade of breast cancer: I grade 2 cases, II grade 40 cases, III grade 18 cases. According to the growth pattern(in number of acinar cells under the microscope for the number of basis) is divided into three types: the cell hyperplasia in 24 cases, all cells and stromal hyperplasia in 24 cases, 12 cases with interstitial hyperplasia. Inclusion criteria:(1) patients with no history of radiotherapy and chemotherapy.(2)patients were informed consent according to the "medical institution regulations".Hematoxylin-eosin staining: the specimens were fixed, embedded in paraffin, serially sectioned in 5?m thick, conventional HE staining, mounted. The performance of each group was observed under light microscope.Immunohistochemistry(SP) staining: according to the routine method, the immunohistochemical stainings for HER2, VEGF, Ki-67, TOPO?? respectively in normal breast, breast hyperplasia, fibroadenoma and breast cancer were done by the corresponding antibody solutions. These specimens were observed under the microscope and took photoes. The positive expression criteria:(1) the staining for HER2 was mainly stained in cell membrane and a small amounts of cytoplasm. No staining or ?10% of the cells whose cell membrane were incompletely, slightly stained are(-); > 10% of the cells were slightly, incompletely stained in cell membrane(+); > 10% of the cells were incompletely and moderately stained in cell membrane or ?10% of the cells showed a strong and complete membrane stained(++); > 10% of the cells showed strong complete membrane stained(+++).(-),(+) And(+) were negative,(+++) was positive.(2)the staining for VEGF was mainly stained in the cytoplasm and a small amount of nuclear. Judge the results by semi-quantitative integration,referring to the scoring criteria of Shimizu et. The intensity of staining was determined: Score of 0: No staining; Score of 1: slight staining; Score of 2: moderate staining; Score of 3: strong staining. The proportion of positive cells was determined:Score of 0: positive cells account for <10%; Score of 1: positive cells account for 10%-30%; Score of 2: positive cells account for 30%-70%;Score of 3:positive cells account for >70%. Finally, the stain intensity score and percentage of positive cells were scored as the final result, ?3 divided into positive(score of 3 and 4 were weakly positive: +; score of 6 was positive: +; score of 9 was strongly positive: +++).(3) Ki67 was stained in nuclear,the results were determined according to the ratio of the positive cells accounted for all cells : negative(-): positive cells?5%; weakly positive(+): positive cells5%-25%; positive(++): positive cells 25%-50%; strongly positive(+++): positive cells > 50%.(4) TOPOII was stained in nuclear,the results were determined according to the ratio of the positive cells accounted for all cells : negative(-): stained cells <10%; positive: stained cells ?10%. Further positive criteria: positive(+): positive cells 10%-25%; positive(++): positive cells 25%-50%; positive(+++): positive cells> 50%.Statistical analysis was performed with SPSS 13.0 software and calculated by chi-square or Fisher exact test, P<0.05 was considered as statistically significant.Results:1 HE staining showed that normal breast tissue consisted primarily of the glandular lobule, glandular lobule could be divided into acinar cells and stroma, according to the position, stroma was further divided into intralobular stroma and interlobular stroma. Breast hyperplasia(sclerosing adenosis) were mainly manifested as the significantly increased number of acini, the less uniformed size and staining of acinar cell nucleus and the varying degrees of hyperplasia in intralobular and interlobular stroma, of those the intralobular stroma hyperplasia was more obvious. Breast fibroadenoma was characterized by intralobular stroma hyperplasia, mucoid degeneration could be seen, the number of acini becomed obviously less, acinar cavity was squeezed into slit, acinar cells had vacuolar degeneration and interlobular stroma was pressed into a thin layer. The histological features of breast cancer were varied, according to their cytological features, they were divided into three types, which were the type of majoring in acinar cell hyperplasia, both acinar cell and stroma hyperplasia and majoring in stroma hyperplasia.2 Immunohistochemical results2.1 The positive expression of HER2 in normal breast tissue, breast hyperplasia(sclerosing adenosis) and breast fibroadenoma was mainly seen in acinar cells, which presented as varying amounts(less than 10%, 50% and 30%, respectively) of the acinar cell membrane were incompletely and slightly stained and all of them were negative. The overall positive rate of HER2 in breast invasive ductal carcinoma was 45%. Among them, in the cases based on cell proliferation, positive cells could be seen gathered into groups, the staining ratio of cell membrane was very high, cell membrane was strongly and completely stained and part of the cytoplasm were also visible shades of staining, positive rate was 75%;In the cases based on both cell and stroma hyperplasia, the number of positive cells was less than cell hyperplasia and the staining was lighter, the positive rate was 37.5%; In the cases based on stroma hyperplasia, the number of positive cells was less, the staining of cell membrane was slight to moderate and most membrane were incompletely stained, all of the cases were negative. The positive expression was mainly found in the acinar cells, and a small amount of positive expression was observed in the stroma. HER2 positive cases with lymph node metastasis accounted for 62.96%(17/27), HER2 negative cases with lymph node metastasis accounted for 42.42%(14/33).2.2 The positive expression of VEGF in acinar cells of normal breast tissue and breast fibroadenoma presented as mainly in cell nuclear staining, cytoplasm was lighter; in the cytoplasm of acinar cells of breast hyperplasia(sclerosing adenosis), the expression of VEGF was more obvious. The expression of VEGF in these three tissues was negative. In the invasive ductal breast carcinoma, the positive expression of VEGF was observed in both acinar cells and stroma, and the acinar cell cytoplasm of the VEGF was strongly stained, the positive rate was 91.67%; In stroma, there were a large number of densely packed cells with different shapes and sizes, and the positive rate was 66.67%. In the case of cell proliferation, the positive rates of acinar cells and stroma were 95.83% and 66.67%, respectively; In the case of both cell and stroma hyperplasia, about 1/3 cases were observed a large number of lymphocytes, and the positive rates of the acinar cell and stroma were 91.67% and 50%, respectively; In the cases of stroma hyperplasia, a large number of blood vessels and local lymphocytes were found be positive, and the positive rates of acinar cells and stroma were 83.33% and 100%, respectively. In the cases of positive expression of VEGF in acinar cells, the rate of lymph node metastasis was 54.54%(30/55), and the rate of lymph node metastasis was 20%(1/5) in the cases with negative expression. In the cases of positive expression of VEGF in stroma, the rate of lymph node metastasis was 45%(18/40), and the rate of lymph node metastasis was 65%(13/20) in the cases with negative expression.2.3 Ki67-positive and TOPO??-positive cells were not found in the acinar cells and stroma of normal breast tissue, while a single positive cell might be seen occasionally within that of breast hyperplasia(sclerosing adenosis) and breast fibroadenoma. Ki67 and TOPO?? were mainly expressed in acinar cells of breast invasive ductal carcinoma, and small amounts were expressed in the stroma. The total positive rates were 78.33% and 45%, respectively. As pathological grade increased, the positive rate increasesd at the same time. The positive rates of Ki67 and TOPO?? in the cases of cell hyperplasia were 83.3% and 66.67%, respectively; In the cases of both cell and stroma hyperplasia, the positive rates were 79.17% and 37.5%, respectively; In th cases of stroma hyperplasia, the positive rates were 66.67% and 16.67%, respectively. The rate of Ki67 positive cases with lymph node metastasis was 59.57%(28/47)and the rate of negative cases with lymph node metastasis was 23.07%(3/13). The rate of TOPO?? positive cases with lymph node metastasis was 66.67%(18/27)and the rate of TOPO?? negative cases with lymph node metastasis was 39.39%(13/33).3 Statistical analysis of Immunohistochemical results:The positive expression of HER2 was related to the growth pattern of breast cancer cells(P=0.000), but it had no relationship with lymph node metastasis and pathologic grade.The positive expression of TOPO?? was related to lymph node metastasis(P=0.026), pathologic grade(P=0.009) and the growth pattern of breast cancer cells(P=0.011).The positive expression of Ki67 was correlated to lymph node metastasis(P=0.02), but it had no relationship with pathologic grade and the growth pattern of breast cancer cells.The positive expression of VEGF in stroma was related to the growth pattern(P=0.011), but it had no relationship with lymph node metastasis and pathologic grade.The positive expression of VEGF in acinar cells had no relationship with lymph node metastasis, pathologic grade and the growth pattern of breast cancer cells.Conclusions:1 The expression of HER2, Ki67, Topo II alpha were negative both in acinar cells and in stroma in normal breast tissue, breast hyperplasia(sclerosing adenosis) and fibroadenoma. Whereas, the expression of VEGF were positive in acinar cells, but negative in stroma in normal breast tissue, breast hyperplasia(sclerosing adenosis) and fibroadenoma.2 Positive expression of HER2, VEGF, Ki67 and Topo II alpha were seen in breast infiltrating ductal carcinoma. HER2 only expressed in acinar cells, mainly expressed in cell membrane, few in cytoplasm; Ki67 and Topo II alpha positive expression were all located in nuclear, mainly in acinar cells, few in stroma. VEGF positive expression were all located in cytoplasm, both in acinar cells and stroma.3 Positive expression of HER2, stromal VEGF, Ki67 and TOPOII alpha in breast hints malignant disease.4 The expression of VEGF and HER2 in breast invasive ductal carcinoma was related to the way of cell growth, Ki67 expression was related to lymph node metastasis, TOPO?? expression was correlated with lymph node metastasis, pathological stage and cell growth mode.