骨形态发生蛋白9体外调节乳腺癌细胞与骨髓间充质干细胞的相互作用

 目的 观察骨形态发生蛋白9（BMP9）在共培养条件下对HS-5成骨分化及MDA-MB-231转移相关因子表达的影响。方法 采用Transwell小室间接共培养MDA-MB-231细胞与HS-5细胞，加入BMP9条件培养基，使用化学发光法、细胞化学染色法及茜素红S染色法分别检测HS-5细胞碱性磷酸酶（ALP）活性、ALP表达及细胞钙盐沉积的变化；应用RT-PCR及Western blot观察HS-5细胞骨钙素（OCN）、MDA-MB-231细胞甲状旁腺激素相关蛋白（PTH-rP）及白细胞介素6(IL-6)mRNA和蛋白表达的变化。结果 HS-5的ALP活性升高，并呈时间依赖性，第9天达最高，随后降低；ALP细胞化学染色结果进一步证实这一点；HS-5细胞钙盐沉积增多、OCN mRNA和蛋白的表达升高（P<0.05）；MDA-MB-231转移相关的PTH-rP及IL-6 mRNA和蛋白的表达下降（P<0.05）。结论 BMP9能够调节乳腺癌细胞与骨髓间充质干细胞的相互作用。

Bone morphogenetic protein 9 regulates the cross-talk between breast cancer cells and bone marrow mesenchymal stem cells

Objective To investigate the effect of bone morphogenetic protein 9(BMP9) on osteogenic differentiation of HS-5 cells and the expressions of metastasis-related factors of MDA-MB-231 cells. Methods The co-culture system with Transwell chamber was exposed to the conditioned media containing BMP9. Then the changes in the activity and expression of alkaline phosphatase (ALP) and calcium deposition in HS-5 cells were observed by chemiluminescence assay、cytochemical staining and Alizarin Red S staining respectively. RT-PCR and Western blot were carried out to detect the mRNA and protein expressions of osteocalcin(OCN) in HS-5 cells、parathyroid hormone-related protein (PTH-rP) and interleukin-6(IL-6) in MDA-MB-231 cells. Results The ALP activity was increased in a time-dependent manner and reached the peak on the ninth day in HS-5 cells，and the cytochemical staining further confirmed that. Compared with the control group,BMP9 increased calcium deposition and the mRNA and protein expressions of OCN in HS-5 cells（P<0.05）,but decreased the mRNA and protein expressions of PTH-rP and IL-6 in MDA-MB-231 cells（P<0.05）.Conclusion BMP9 could regulate the cross-talk between breast cancer cells and bone marrow mesenchymal stem cells.