摘 要 目的: 应用差异凝胶电泳分析骨肉瘤多药耐药细胞株与亲本细胞株蛋白质表达的差别，筛查骨肉瘤细胞多药耐药相关蛋白。方法：以小剂量多柔比星（doxorubicin, DXR）诱导人骨肉瘤细胞株MG63，建立骨肉瘤多药耐药细胞株MG63/DXR100。以差异凝胶电泳分离两组细胞中的全部蛋白质，应用图像扫描和DeCyder软件分析差异（表达增加或减少>30%）表达的蛋白质点，对其进行质谱鉴定，并在Mascot 数据库中检索。结果：共检测到明显差异表达的蛋白质点30个，选择其中18个点进行质谱鉴定，有5个蛋白质点鉴定成功，它们分别是与肿瘤细胞转移相关的基质金属蛋白酶1（matrix metalloproteinases 1, MMP1）、具有解毒功能的乙醇脱氢酶6（alcohol dehydrogenase 6, ADH6）、属于抑癌基因的FERM域结合蛋白3（FERM domain containing protein 3, FRMD3），以及两个分别由128和300个氨基酸残基构成的未知蛋白。MMP1、ADH6和2种未知蛋白在耐药细胞表达显著高于非耐药细胞组，而FRMD3表达显著显著低于非耐药细胞。结论：通过差异凝胶电泳筛查到，MMP1、ADH6、FRMD3 及2种未知蛋白质在骨肉瘤细胞的多药耐药细胞和非耐药细胞中差异表达，它们可能参与骨肉瘤细胞的多药耐药机制。

Abstract Objective: To analyze the differential protein expression between multidrugresistant human osteosarcoma MG63/DXR100 and its parental cell line by differential ingel electrophoresis, so as to lay a foundation for exploring the mechanism of multidrug resistance(MDR) of osteosarcoma. Methods: Multidrugresistant clone of human osteosarcoma MG63 was established by stepwise exposure to increasing doses of doxorubicin(DXR). The total proteins of the above two cell lines were separated with twodimensional electrophoresis. The proteins of differential expression (increased or decreased by more than 30%) were analyzed with image scanning and DeCyder software. Then they were identified in MALDITOF Pro and Mascot database. Results: Thirty proteins with differential expression were identified by proteomic analysis, and 18 of them were further identified by MALDITOF Pro. Five protein spots were successfully identified: matrix metalloproteinases 1 (MMP1) related with tumor cell metastasis, alcohol dehydrogenase (ADH6) with function of detoxication, FERM domain containing protein 3 (FRMD3) which belongs to antioncogenes and two agnoproteins composed of 128 and 300 amino acid residues. MMP1, ADH6 and the two agnoproteins were overexpressed in MG63/DXR100 group, and FRMD3 expression was lower than that in the MG63 group. Conclusion: Five proteins including MMP1, ADH6, FRMD3 and two agnoproteins have been found abnormally expressed in MDR osteosarcoma cells by differential ingel electrophoresis; they might participate in MDR of osteosarcoma.

国家自然科学基金资助项目（No.30772210）