The Expression Of Stromal CD10, Ezrin And Nek2 Associated With The Development And Progression Of Breast Carcinoma

Part1ObjectiveTo evaluate the expression of stromal CD 10 and Ezrin in ductal carcinoma in situ(DCIS) and invasive ductal carcinoma (IDC)of breast cancer, to observe the relationship of their expression with breast cancer pathological features and prognosis and to examine their role in tumor progression.MethodsImmunohistochemistry was used to examine the expression ofstromal CD 10 and Ezrin in 60 specimens of DCIS (30 specimens of low/intermediate grade DCIS and 30 specimens of high grade DCIS) and 60 specimens of IDC (30 specimens ofⅠ/Ⅱgrade and 30 specimens ofⅢgrade). The breast cancer tissue which expressed stromal CD 10 and Ezrin was selected as a positive control,and in the negative control group the stromal CD 10 and Ezrin antibody were replaced by PBS.Results(1)In DCIS the expression of stromal CD 10 was negative in 36 cases, weak in 18 cases,strong in 6 cases, whereas in IDC the corresponding part was 25 cases,18 cases,17 cases respectively, it was preferentially expressed in IDC compared to DCIS(z=-2.465,P=0.014). There was no correlation between the expression of stromal CD10 and tumor grade(z=-0.115,P>0.05),tumor size(z=-0.892,P>0.05),ER (z=-0.647,P>0.05)and PR(z=-0.213,P>0.05) status,and lymph node metastases in IDC(z=-0.712,P>0.05).(2)In DCIS Ezrin localized at apical was seen in 15 cases, at membrane was seen in 13 cases and in cytoplasm was seen in 21 cases,whereas in IDC the corresponding part was 5,6,38 respectively, its localization was significantly different in this two groups (x2=12.477,P=0.002). There was no correlation between the expression of Ezrin and tumor grade(x2=0.733,P> 0.05),tumor size(x2=0.314,P>0.05),ER (x2=3.464,P>0.05)and PR(x2=2.195,P> 0.05) status,and lymph node metastases in IDC(x2=3.281,P>0.05).(3)The expression of CD 10 and switch of Ezrin localization were positive correlated with disease free survival(χ2=4.062,P<0.05;χ2=4.398 P<0.05).ConclusionsCompared to DCIS stromal CD 10 was preferentially expressed in IDC,and the localization of Ezrin was significantly different in this two groups.The increase of expression of stromal CD 10 and the switch of Ezrin localization might participate in the process of invasion of DCIS, and were associated with poor prognosis. Part2ObjectiveTo investigate the expression and location of Nek2 during development and progression of breast cancer, as well as the relationship between changes of Nek2 and centrosome defects.To evaluate the role of Nek2 in progression of breast cancer.MethodsThe multi-step simple linear progression model of breast cancer was composed of MCF10A, MCF10AT, MCF10DCIS.com and MCF10Cal.DNA ploidy was analyzed by flow cytometry. The expression level of Nek2 was evaluated by western blot. Immunofluorescence was used to investigate the location of Nek2 in cells and abnormal centrosome.Results1.Analysis of DNA ploidy:The DNA index of MCF10AT was 1.0, the DNA index of MCF10DCIS.com was 1.2, the DNA index of MCF10Cal was 1.5.2.The percentage of abnormal centrosome in the four cell lines was 1%,3%,5.6%,8%respectively.3.The expression level of Nek2 in MCF10DCIS.com cell was higher than in MCF10AT, highest in MCF10Cal and lowest in MCF10A.Nek2 was expressed in cytoplasm and nucelus in all cell lines.Conc lusions1.MCF10A and MCF10AT were diploidy, MCF10DCIS.com and MCF10Cal were heteroploidy.The DNA ploidy was stable from hyperplasia to atypical hyperplasia,but was changed from atypical hyperplasia to ductal carcinomas in situ.2.The percentage of abnormal centrosome were different in four cell lines.The abnormal centrosome percentages in MCF10DCIS.com and MCF10Cal were higher than that in MCF10A,the percentage in MCF10Cal was higher than that in MCF10AT,but it was the same in MCF10DCIS.com and MCF10Cal cell,and so did between MCF10DCIS.com and MCF10AT, MCF10AT and MCF10A cell. 3. The increase expression of Nek2 in four cell lines explained it's role in promote breast carcinoma development and progression.4.The changes of DNA ploidy, abnormal centrosome percentage and expression of Nek2 support the multi-step simple linear progression model of breast carcinoma.