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目的:比较蛋白酶体抑制剂PS-341处理多发性骨髓瘤细胞U266前后蛋白质组的差异,探究PS-341潜在的药物靶点,为多发性骨髓瘤的临床治疗提供理论依据。方法:用蛋白酶体抑制剂PS-341处理骨髓瘤细胞U266,应用双向凝胶电泳技术分离PS-341处理前后的U266细胞的蛋白质,Im-ageMaster 2D Platinum图像分析软件识别药物处理前后U266细胞的差异表达蛋白质点,基质辅助激光解吸电离飞行时间质谱(MALDI-TOF-MS)鉴定差异表达的蛋白质。Western印迹法检测差异蛋白质BAG-2在药物处理前后U266细胞中的表达水平。结果:建立了PS-341处理前后U266细胞蛋白质的双向凝胶电泳图谱,找到55个差异表达的蛋白质点,鉴定了31个差异表达的蛋白质,有27个蛋白质在PS-341处理后下调。Western印迹分析证实BAG-2在药物处理前后U266细胞中的表达水平存在差异。结论:处理后下调的一些蛋白可能是蛋白酶体抑制剂PS-341潜在的药物靶标。
OBJECTIVE: To compare proteasome differences between proteasome inhibitor PS-341 before and after treatment with multiple myeloma cells U266 and explore the potential drug target of PS-341 to provide a theoretical basis for the clinical treatment of multiple myeloma. Methods: The myeloma cell line U266 was treated with proteasome inhibitor PS-341. The protein of U266 cells before and after treatment with PS-341 was separated by two-dimensional gel electrophoresis. The difference of U266 cells before and after drug treatment was identified by Im-ageMaster 2D Platinum image analysis software Protein spots were identified and differentially expressed proteins identified by matrix-assisted laser desorption / ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Western blotting was used to detect the expression level of BAG-2 protein in U266 cells before and after drug treatment. Results: Two-dimensional gel electrophoresis of proteins of U266 cells was established before and after PS-341 treatment. 55 differentially expressed protein spots were identified, 31 differentially expressed proteins were identified, and 27 proteins were downregulated after PS-341 treatment. Western blot analysis confirmed that there was a difference in the expression level of BAG-2 in U266 cells before and after drug treatment. Conclusion: Some proteins down-regulated may be potential drug target of proteasome inhibitor PS-341.