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《Journal of Third Military Medical University》 2012-01
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Biological characteristics of fibroblasts primarily cultured form breast precancerosis

Sun Yan1,2,Peng Qiongle1,2,Wang Liyang1,2,Zhao Liuyang1,2,Yuan Quan1,Li Jun1,Liu Manran1,2(1Key Laboratory of Diagnostic Medicine of Ministry of Education,2Department of Cell Biology and Genetics,College of Basic Medical Sciences,Chongqing Medical University,Chongqing,400016,China)  
Objective To investigate the biological characteristics of breast primary atypical intraductal hyperplasia fibroblasts(AFs) in tumorigenesis of breast cancer,normal mammary fibroblasts(NFs) and the breast primary cancer-associated fibroblasts(CAFs).Methods The primary human breast fibroblasts were isolated by collagenase Ⅰ digestion from normal mammary tissue,atypical intraductal hyperplasia and mammary tumor.The purity of these fibroblasts was identified by immunofluorescent staining using the specific first antibody to fibronectin,the fibroblast biomarker.The proliferation and cell cycle for each kind of fibroblasts was detected by MTT assay and flow cytometry.The expression of cyclin D1,p53 and p21waf1 in the fibroblasts was detected by Western blotting.Results More than 95% of the isolated fibroblasts were positive to fibronectin.Comparing with NFs(8.24±2.47),AFs(16.94±4.77) had higher growth rate and more cells at S-phase of cell cycle(P0.05).However,there were lower growth rate and lesser S-phase cells(P0.05) when compared these parameters in AFs with that in CAFs(33.31±7.90).Similar changes were found on the expression of cyclin D1,p21waf1 and p53 among AFs,NFs and CAFs.Conclusion Different biological characteristics of AFs with NFs and CAFs indicate that AFs are on the way of transformation of NFs to CAFs in the precancerous lesion of breast cancer.
【Fund】: 国家自然科学基金主任基金(30940096);国家自然科学基金(81072147);; 重庆市科委自然科学基金(CSTC2010BB5099);; 重庆医科大学校重点基金(XBZD201006);重庆医科大学优秀留学归国人员启动基金(0200101189)~~
【CateGory Index】: R737.9
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