目的:分离培养和鉴定乳腺癌相关成纤维细胞及其与之配对的正常乳腺成纤维细胞,并初步探讨两者生物学性状的差异。方法:用胶原酶消化法获得原代乳腺癌相关成纤维细胞及其与之配对的正常乳腺成纤维细胞,通过显微镜观察细胞形态,免疫荧光观测上皮细胞和成纤维细胞标记,M1rr及Western检测两者之间生物学性状差异。结果:分离得到乳腺癌相关成纤维细胞及其与之配对的正常乳腺成纤维细胞,两者均高表达Vimentin低表达cytokeratin,且两者在形态及生物学性状有一定差异。结论:与配对成纤维细胞相比.乳腺癌相关成纤维细胞高表达α-SMA,且增殖能力增强。
Objective The current study was to isolate and identify breast cancer associated fibroblasts and their paired normal breast fibroblasts, and also to explore their biological differences. Methods Primary breast cancer associated fibroblasts and their paired normal breast fibroblasts were isolated from primary breast tumors and breast tissue by collagenase digestion. Morphology was observed by microscope. Cell marker was studied by immunofluorescence staining. Cell proliferation and protein level was separately evaluated by MTT and western blot. Results The protein level of vimentin was highly expressed, however, the protein level of cytokeratin was decreased in the isolated breast cancer associated fibroblasts and their paired normal breast fibroblasts. The two cells showed different features in morphology and proliferation. Conclusion The level of α-SMA was highly expressed, and the cell proliferation was enhanced in the breast cancer associated fibroblasts.