骨唾液酸蛋白的表达对骨肉瘤细胞侵袭转移特性的影响.docVIP

骨唾液酸蛋白的表达对骨肉瘤细胞侵袭转移特性的影响 作者：黄涛，陈安民，郭风劲，张衣北，廖晖 【摘要】 目的 构建人骨唾液酸蛋白（BSP）正反义真核表达载体,研究其对骨肉瘤细胞侵袭转移特性的影响。方法 以PCR的方法扩增人BSP的开放阅读框序列，与载体pIRES2EGFP相连构成重组正反义表达质粒。以脂质体介导转染入MG63细胞，通过Western blot检测细胞中蛋白的表达。以重组基底膜侵袭实验和划痕实验测定对骨肉瘤细胞侵袭力的影响。结果 成功构建了人BSP正反义核酸真核表达载体，转染后有效表达hBSP，与对照组相比，正义载体转染后使MG63细胞中hBSP蛋白表达水平升高，侵袭力增强；反义载体转染后使MG63细胞中hBSP蛋白表达水平降低，侵袭力受到抑制。结论 BSP表达的改变可能在骨肉瘤的浸润转移过程中起重要作用，BSP表达下调可以抑制骨肉瘤的侵袭能力。 【关键词】 骨唾液酸蛋白基因；基因表达；肿瘤转移；骨肉瘤 　　Expression of BSP Influence on Behavior of Invasion and Metastasis of Osteosarcoma　 　　Abstract：Objective To construct sense and antisense human bone sialoprotein (BSP) eukaryotic expression vector and observe influence of its expression in human osteosarcoma cell line MG63 cells about invasion and metastasis of osteosarcoma.Methods An open reading frame of human BSP was amplified by PCR method and was reconstructed 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 lipofec tamine.The expression of BSP on the cells was measured by Western blotting.Transwellchamber methodand scarification was used to evaluate influence of BSP expression on osteosarcoma cell invasion and metastasis.Results The sense and antisense hBSP eukaryotic vector was successfully constructed and transfected. Compared with control cells,expression of BSP increased in the cells by sense hBSP eukaryotic vector transfected and invasive power of these cells is enhance,but expression of BSP decreased in the cells by antisense hBSP eukaryotic vector transfected and invasive power of these cells is been inhabited.Conclusion The change of BSP expression may play an important role in osteosarcoma metastasis.The downregulation of BSP expression can inhibit abilities of osteosarcoma cell invasion and metastasis. 　　Key words：Bone sialoprotein gene; Gene expression; Neoplasm metastasis; Osteos