BMP-4通过Notch信号增强乳腺癌细胞上皮间充质转化和肿瘤干细胞特性的研究

BMP-4 enhances epithelial-mesenchymal transition and cancer stem cell properties in breast cancer cells via Notch signal

目的探讨骨形态发生蛋白-4(BMP-4)通过Notch信号增强乳腺癌细胞上皮间充质转化(EMT)和肿瘤干细胞特性。方法采用Dulbecco改良Eagle培养基F-12培养MCF-10A细胞,细胞保存亚融合状态,用0.05%胰蛋白酶/EDTA传代,使用4~8代细胞。人骨髓源性间充质干细胞保存于间充质干细胞培养基中,采用第3~6代人骨髓间充质干细胞。所有细胞在含有5%的CO2的37℃加湿培养箱中保存,使用倒置显微镜观察细胞的显微图像。结果 BMP-4增强乳腺上皮细胞系和其EMT及干细胞特性,BMP-4增加了EMT生物标志物表达,激活了细胞中的Notch信号,提高了未转化乳腺上皮细胞系MCF-10A的球体形成效率、集落形成效率和肿瘤干细胞标志物的表达路线。结论在乳腺上皮细胞和乳腺癌细胞中,BMP-4通过激活Notch信号,来增加EMT和干细胞特性。

Objective To study the performance of bone morphogenetic protein-4( BMP-4) in enhancing epithelial-mesenchymal transition( EMT) and cancer stem cell properties in breast cancer cells via Notch signal. Methods MCF-10 A cells were cultured in Dulbecco modified Eagle medium F-12. The cells were subcultured with 0. 05% trypsin/EDTA and 4 to 8 passage cells were used. Human bone marrow-derived mesenchymal stem cells were preserved in mesenchymal stem cell culture medium. Passage 3 to 6 of human bone marrow mesenchymal stem cells were used in all experiments. All cells were stored in a humidified incubator containing 5% CO2 at 37℃. An inverted microscope was used to observe microscope image of cells. Results BMP-4 enhanced EMT and stem cell properties of breast epithelial cell lines and breast cancer cell lines. Moreover,BMP-4 increased the expression of EMT biomarkers,activated the Notch signal in cells and improved the sphere forming efficiency of the non-transformed mammary epithelial cell line MCF-10 A,colony formation efficiency and the expression route of cancer stem cell markers. Conclusion BMP-4 can increase EMT and stem cell characteristics by activating Notch signal in breast epithelial cells and breast cancer cells.