| Objective Metaplastic breast carcinoma(MBC) is a rare heterogeneous group of primary breast malignancies, with more frequent triple-negative subtype and poor outcomes. So far no prognostic markers for the tumor are validated yet and data from the Chinese population is scarce. The first part of the study is undertaken to evaluate its clinicopathologic characteristics, responses to therapeutic regimes, prognosis and patients whether benefit from EGFR targeted therapy or not in a large cohort of patients from a major Chinese cancer center. IL-17(Interleukin-17) is a pro-inflammatory cytokine that mainly produced by activated CD4~+T cells(also known as Th17 cells).The study have showed that IL-17 might facilitate development of cancer by fostering angiogenesis through promote VEGF production from cancer cells. While, there are some studies have indicated that the increased number of Th17 cells was associated with good prognosis of patients. The second part of this study aims to investigate the location of IL-17~+cells in invasive ductal carcinoma not otherwise specified(IDC-NOS) and medullary carcinoma(MC), its relationship with CD4~+T cells, VEGF, clinicopathologic characteristics and prognosis were analyzed. We aim to analyse the location of IL-17~+cell in breast cancer microenvironment and study IL-17 how to affect the development of tumor.Methods Ninety cases of MBC were identified in the Department of Breast Cancer Pathology and Research Laboratory, Tianjin Medical University Cancer Hospital between January 2000 and September 2014, and we compared to the cases of 1090 patients with invasive ductal carcinoma not otherwise specified(IDC-NOS) from the same time period. By immunohistochemical staining(IHC) and fluorescence in situ hybridization(FISH), the expression of ER, PR, HER2, Ki-67,p53,CK5/6 and EGFR were analyzed and we studied the relationship between clinicopathologic characteristics and prognosis in patients with MBC.114 cases of IDC-NOS with tumor infiltrating lymphocytes(>40%) from 521 cases of IDC-NOS were identified in our hospital between January 2008 and April 2009, and we compared to the cases of 70 patients with medullary carcinoma between January 2006 and December 2010. By immunohistochemical staining, the location of IL-17~+cells,CD4~+T cells and the expression of VEGF were determined in IDC-NOS and medullary carcinoma. The relationship between IL-17~+cells and CD4~+T cells, the expression of VEGF and with the clinicopathologic characteristics, which include the prognosis of patients were determined.Results 1. Compared to patients with IDC-NOS or TN-IDC, patients with MBC displayed larger tumor size(P =0.000, P=0.013), less frequent lymph node metastasis(P =0.000, P=0.000) and higher rate of local recurrence or distant metastasis(P=0.001, P=0.044) and a greater proportion of MBCs are TNBCs than tumors in the IDC-NOS group(P=0.000). 2. Fewer patients in the MBC group received MRM(P=0.000, P=0.000), radiotherapy(P=0.000, P=0.001), endocrine therapy(P=0.000) and anti-HER2 therapy(P=0.000) than those in the TN-IDC and/or IDC-NOS group. 3. The most common MBC subtype was spindle cell carcinoma(34.4%), followed by squamous cell carcinoma(31.1%) and MBC with mesenchymal differentiation(24.5%). Fibromatosis-like subtype(4.4%) was the least common in this cohort of patients. 4. In MBC patients, two cases were classified as luminal A type(2.2%), 17 cases were luminal B type(18.9%), 7 cases were HER2-overexpression type(7.8%), and 64 cases were triple-negative type(71.1%). EGFR overexpression was identified in 52(57.8%) cases. Forty-seven of 52 cases with EGFR overexpression were submitted for FISH analysis, and 14 of them(29.8%) demonstrated EGFR gene amplification. 5. In univariate analysis of MBC, LNM(P=0.000), advanced stage at diagnosis(P=0.047), EGFR overexpression(P=0.007), and high Ki-67 labeling(P=0.026) of tumors were the significant predictive factors for reduced DFS, among which only LNM(P=0.008) was significant in multivariate analysis. For OS, tumor lymph node metastasis(P=0.000),EGFR overexpression(P=0.049), and gene amplification(P=0.002) were the significant predictive factors in univariate analysis, among which lymph node metastasis and EGFR gene amplification were significant predictive factors using multivariate analysis(P=0.001 and 0.022). 6. Compared to patients with medullary carcinoma, patients with IDC-NOS displayed some characteristics, such as more frequent lymph node metastasis(P=0.002) and triple-negative subtype(P=0.003). 7. The distribution of IL-17~+ cells and CD4~+T cells in breast cancer: Compared with the control group, the density of IL-17~+cells(including in intratumor, peritumoral stroma and distant stroma) was significantly higher in IDC-NOS tissue(P=0.001, P<0.001, P<0.001, P<0.001). The density of CD4~+T cells was significantly higher in distant stroma of medullary carcinoma than IDC-NOS group(P=0.016). 8. The correlation analysis between IL-17~+ cells and CD4~+T cells in breast cancer: In IDC-NOS group, we found a significant correlation between the number of IL-17~+ cells and CD4~+T cells(r=0.222, P=0.017) in distant stromal, but there was no significant correlation in intratumor and peritumoral stroma. While, there was a positive correlation between the number of IL-17~+cells and CD4~+T cells in intratumor of medullary carcinoma(r=0.236, P=0.049). 9. In IDC-NOS group, we identified that the expression of VEGF was positively correlated with the number of IL-17~+cells in different parts of tumor tissue(P?0.001). 10. In IDC-NOS, the number of IL-17~+ cells had a significant positive correlation with ER(P=0.030) only. And the expression of VEGF had no significant correlation with clinicopathologic characteristics, including age, TNM stage, the expression of receptor, et al. 11. In univariate analysis, LNM(P=0.011), stage at diagnosis(P=0.013) were the significant predictive factors for DFS, For OS, the status of ER(P=0.017) and triple-negative subtype(P=0.012) were the significant predictive factors in univariate analysis. Next, we found that the higher number of IL-17~+ cells in intratumor and peritumoral stroma was the significant predictive factor for reduced OS(P=0.015, P =0.037).Conclusions 1. The tumors of MBC presented with lager size, lower rate of lymph node metastasis, and demonstrated more frequent local recurrence/distant metastasis in follow-up than the cases of IDC-NOS. 2. On univariate analysis of MBC cases, lymph node metastasis, advanced clinical stage at diagnosis, high tumor proliferation rate assessed by Ki-67 labeling, and EGFR overexpression of tumors were associated significantly with reduced DFS, while decreased OS was associated significantly with lymph node metastasis and EGFR overexpression/gene amplification. On multivariate analysis status of lymph node was proven to be an independent predictor for DFS, and status of lymph node and EGFR gene amplification to be the independent predictors for OS. 3. The study indicates that MBC is an aggressive type of breast cancer with poor prognosis, and efficient comprehensive therapeutic regimens are to be identified. And many MBC patients showed EGFR overexpression and/or gene amplification, new therapy targeting EGFR in tumors is worthy of further clinical research. 4. In general, IDC-NOS presented with a higher rate of lymph node metastasis, poor prognosis and demonstrated more frequent triple-negative subtype than medullary carcinoma. 5. The density of IL-17~+cells(including in intratumor, peritumoral stroma and distant stroma) was significantly higher in IDC-NOS tissue than control group, indicating that the higher expression of IL-17 may be associated with the degree of malignancy. 6. IL-17 may facilitate development of breast cancer by promoting VEGF production from cancer cells and fostering angiogenesis. 7. The IL-17~+ cells infiltrating in various parts of IDC-NOS tissue were associated with the prognosis of the patients with different meanings, and Th17 cell mainly distributed in distant stroma of IDC-NOS tissue, while in medullary carcinoma it mainly distributed in intratumor, suggest that the different distribution of Th17 cell in breast cancer microenvironment may be one of the factors affecting the prognosis of the patients. 8. The function of IL-17 and Th17 cells in tumor progression and prognosis is controversial. Although there are certain intersections between IL-17 and Th17 cells,they may have different effects in tumor progression and IL-17 can not be equivalent to Th17 cells. |
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