| Objective:Phyllodes tumors of the breast are rare neoplasms that account for <1% of all mammary tumors and 2-3% of fibro-epithelial neoplasms of the breast.We evaluated the clinicopathological characteristics of a cohort of 246 Chinese patients in relation to the expression of epithelial-to-mesenchymal(EMT)markers in benign,borderline and malignant tumors and the prognostic value of different surgical regimens.And we investigated whether the interactions between mammary epithelial cells and human fibrosarcoma cells affect the morphology,proliferation,and differentiation of epithelial cells in a co-culture system.Methods:After analyzing the clinicopathological parameters of breast cancer patients at the Cancer Hospital of Tianjin Medical University from April 1987 to June 2010,246 patients ranging from 11 to 76 years old with a median age of 46 years were selected for our study.The expression of EMT and mesenchymal stem cell biomarkers were evaluated for all 246 cases by performing IHC and IF.The cases were followed-up until March 1,2016 and ranged from 60 to 345 months with a median duration of102.5 months.Disease-free survival(DFS)was defined from the date of surgery to the date of local relapse and/or distant metastasis.Reappearance of tumor in the treated remnant breast,skin or chest wall was defined as local recurrence.Distant metastasis was considered when the tumor metastasized at a distant site like lungs,bone and/or liver.To investigate the effects of HT1080 on proliferation and differentiation of mammary epithelial cells,we established the co-culture system with indirect contact between HT1080 condition-medium and MCF-10 A to verified relative EMT expression.Results:Of the 246 patients,significant differences existed between three groups according to different surgical managements(breast-conserving surgery,lumpectomy,mastectomy or modified/ radical mastectomy;P < 0.001).The expression ofE-cadherin,Snail,Slug and Twist in epithelial cells of benign PTs was lower than the borderline and malignant PTs,(P=0.025,P=0.001,P<0.001,P=0.044,respectively),no difference was observed for N-cadherin(P=0.146).For the EMT markers highlighting the positive stroma component,Snail(P=0.007)and Slug(P=0.001)increased with the advancing grade of PTs.And co-expression of MSC(GD2 and ALDH1)and EMT markers was observed in 15 cases.Also,nuclear(Slug and Twist)and cytoplasmic(GD2 and ALDH1)staining was observed in these malignant PTs.Based on the Kaplan–Meier and log-rank statistic methods,we observed that the OS,DFS,5-year DFS and 5-year OS was significantly lower in the malignant group in comparison to the borderline and benign groups.Based on univariate analysis,advanced grade(P=0.001),Slug overexpression(P= 0.036)and surgical management(P < 0.01)of PTs were the significant predictive factors for DFS.Only surgical management(P=0.009)was significant in the multivariate analysis.For OS,advanced grade at diagnosis(P < 0.01),history of recent growth(P=0.007),Slug overexpression(P=0.028)and surgical management(P=0.002)of tumors were the significant predictive factors in univariate analysis,whereas,history of recent growth and grade of PTs were significant predictive factors in multivariate analysis(P=0.003 and 0.001).MCF-10A/HT1080 co-cultures exhibited a variety of changes as compared with MCF-10 A monoculture,including(a)morphologic changes,a typical epithelial morphology to a spindle-like,fibroblastic morphology;(b)cytoskeleton re-constitution,scatter arrangement to densely interwoven filaments by F-actin staining;(c)an EMT-like expression profile,suppression of the epithelial marker,E-cadherin,and expression of mesenchymal markers,N-cadherin and vimentin;and expression of EMT markers,Snail,Slug and Twist.(d)level of EMT mRNA improvement by RT-PCR.In contrast to what we observed with TGF-β+ MCF-10 A cells,the stimulatory effect of TGF-β+ on differentiation of mammary epithelial cells was observed after culture of MCF-10 A cells with TGF-β+-conditioned medium. |
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