人骨唾液酸蛋白正反义真核表达载体的构建及在骨肉瘤细胞中的表达

目的 构建人骨唾液酸蛋白（hBSP）正反义真核表达载体，并检测其在骨肉瘤细胞MG-63中的表达。方法 以人骨膜细胞总RNA为模板，采用RT-PCR技术扩增出hBSP全长序列，通过DNA重组技术构建hBSP正反义核酸真核表达载体，并用脂质体介导转染人MG-63细胞，通过RT-PCR及Western blot检测细胞中hBSPmRNA和蛋白的表达。结果 成功构建了hBSP正反义核酸真核表达载体，转染后有效表达hBSP，正义载体转染后使MG-63细胞中hBSPmRNA和蛋白表达水平升高，反义载体转染后使MG-63细胞中hBSPmRNA和蛋白表达水平降低。结论 转染正反义hBSP表达载体导致瘤细胞BSP表达的改变，为进一步实验研究提供了基础。

Construction of Sense and Antisense Human Bone Sialoprotein Eukaryotic Expression Vector and the Expression in Osteosarcoma Cells

Objective To construct sense and antisense human bone sialoprotein(hBSP) eukaryotic expression vector and detect its expression in human osteosarcoma cell line MG-63 cells.Methods Human BSP full-length cDNA was amplified by RT-PCR method from human periosteum and was transfected into the eukaryotic expressive vector pIRES2-EGFP to construct the hBSP sense or antisense expressive vector.The sense and antisense hBSP eukaryotic expression vector was transfected into MG-63 cells with lipofectamine.The expression of hBSP in the cells was detected by RT-PCR and Western blot.Results The sense and antisense hBSP eukaryotic expression vestor was successfully constructed and transfected.The mRNA and protein expression of hBSP was increased in the cells transfected by sense hBSP eukaryotic vector but decreased in those transfected by antisense hBSP eukaryotic vector.Conclusion Transfection of the sense or antisense hBSP eukaryotic expression vector can result in the changes in the expression of BSP in MG-63 cells.