摘要
目的:比较骨肉瘤患者与正常人血清的差异蛋白质,为对骨肉瘤的诊断、治疗、预后的判断等提供依据。方法:应用cleanup试剂盒分别对骨肉瘤患者血清及正常人血清进行浓缩,然后去高丰度蛋白质,2D clean-up kit试剂对蛋白质纯化后进行双向凝胶电泳,银染扫描后用Image Master 2DE软件分析寻找差异蛋白,切除差异蛋白质点酶解后进MALDI TOF/TOF(基质辅助激光解吸电离飞行时间质谱)质谱鉴定,获取肽指纹图谱并查询SwissProt数据库寻找匹配蛋白质。结果:获得重复性较好的双向电泳银染图谱。蛋白质点经质谱分析并检索SwissProt数据库后,发现11个差异蛋白质,其中5个上调,6个下调。结论:骨肉瘤患者血清与正常人血清蛋白质组存在明显差异,角蛋白、生长激素、内质网蛋白29等差异蛋白质有望成为诊断治疗骨肉瘤的分子标记物或者治疗靶点,为进一步对骨肉瘤的研究提供理论依据。
Objective Tocompare the differences of the serum proteomics between patients with osteosarcoma and normal individuals and to identify serum biomarkers that were useful in the diagnosis, treatment and prognosis of osteosarcoma. Methods Apply cleanup kit to concentrate total protein from nine osteosarcoma serum samples and normal serum, then removal of high abundance proteins respectively. And the total protein was purified by 2-DE the clean-up kit reagents. Then dielectrophphoresis was conducted. To find out differential protein by Image Master 2-DE software analysis after silver staining and scanning. Then, MALDI TOF/TOF mass spectrometric identification of differential protein spots was conducted. Get access to the peptide mass fingerprints and query to find out matching proteins in the Swiss-Prot database. Results Dieletrophoresis silver staining map obtained had good repeatability. The mass spectrometry analysis revealed 5 unique proteins increased, and 6 unique protein decreased in osteosarcoma serum samples. Conclusions The results suggest that proteomic analysis can identify protein varianes between the osteosarcoma serum and normal serum, so provide a theoretical basis for the further study of osteosarcom.
出处
《湖南师范大学学报（医学版）》
2018年第1期42-46,共5页
Journal of Hunan Normal University(Medical Sciences)
