骨桥蛋白促MV4-11白血病细胞uPA表达及信号通路研究

Study on osteopontin stimulating expression of urokinase type plasminogen activator and its role in the signaling pathway in MV4-11 leukemic cells

目的初步研究骨桥蛋白(osteopontin,OPN)在MV4-11白血病细胞株促尿激酶纤溶酶原激活物(urokinase type plasminogen activator,u PA)表达的信号通路。方法 1)细胞培养:IMDM培养基培养MV4-11白血病细胞。2)分组:1人重组OPN蛋白(0-0.3)μg/m L处理MV4-11白血病细胞24h组;2 PI3-K抑制剂(0-30)μmo L,LY294002处理MV4-11白血病细胞1 h,再用OPN 0.3μg/m L处理细胞24 h组;3NF-ΚB抑制剂(0-15)μmo L处理MV4-11白血病细胞1 h,再用OPN 0.3μg/m L处理细胞24 h组。3)Western blot法分别检测三组细胞u PA表达。结果人重组OPN蛋白单独处理组,随OPN蛋白的浓度增加,u PA表达增加;预先用LY294002及BAY 11-7082处理组,OPN诱导的u PA表达降低。结论 OPN能促进MV4-11白血病细胞u PA表达;LY294002及BAY 11-7082均阻断了OPN促u PA蛋白表达效应,PI3K及NF-KB是这一信号通路中的关键分子。

Objective To conduct a preliminary study on osteopontin（ OPN） stimulating expression of urokinase type plasminogen activator（ u PA） and its role in the signaling pathway in MV4-11 leukemia cells. Methods MV4-11 cells were cultured in IMDM medium. Then,MV4-11 cells were categorized into three groups,for instance,the OPN group（ treated with 0- 0. 3 μg / m L soluble Recombinant Human Osteopontin for 24h）,the inhibitor of PI3-K group（ pretreated with 0- 30 μmol LY294002 for 1h and then treated with 0. 3 μg / m L soluble Recombinant Human Osteopontin for additional 24 h）,and the inhibitor of NF-ΚB group（ pretreated with 0- 15 μmol BAY 11- 7082 for 1h and then treated with 0. 3 μg / m L soluble Recombinant Human Osteopontin for additional 24 h）. The expression of u PA protein was detected by Western Blot. Results In the OPN group,the expression of u PA protein increased along with the increasing dose of soluble Recombinant Human Osteopontin. In other groups,the expression of u PA protein induced by OPN reduced significantly. Conclusion OPN can enhance the expression of u PA. The inhibitor of PI3-K and NF-ΚB reduced the expression of u PA protein induced by OPN. PI3-K and NF-ΚB are the key molecules in this signaling pathway.