CAJ | 학술논문

目的应用iTRAQ技术观察BMP-2诱导肌源C2C12细胞向成骨细胞分化过程中蛋白质表达组的变化.方法将肌源C2C12细胞接种于BMP-2诱导分化体系中进行分化诱导,提取第7天的分化蛋白以iTRAQ试剂标记后进行质谱检测,分析差异表达的蛋白质,并进行生物信息学分析.结果 iTRAQ试剂标记的BMP-2诱导肌源C2C12分化细胞蛋白质表达谱分析筛选出明显差异表达蛋白质点为23个,表达上调的蛋白质点8个,下调的蛋白质点15个.通过趋势分类发现上述差异蛋白在C2C12细胞成骨分化的各时期(第1至7天)均存在蛋白的差异表达.其中部分上调蛋白在分化早期表达水平升高、部分上调蛋白在分化晚期表达水平升高;同样,部分下调蛋白在分化早期表达水平下降,部分下调蛋白在分化晚期表达水平下降.初步鉴定SERCA3、细胞色素b5、S100A4、ATPase inhibitor、ATPIF1等5个蛋白表达出现动态变化,证实上述蛋白可能与诱导成骨分化机制有关.结论本研究差异蛋白表达趋势的结果显示全程监测C2C12细胞成骨分化的必要性,提示iTRAQ技术是研究细胞分子蛋白改变的有效的蛋白质组学方法,SERCA3、细胞色素b5、S100A4、ATPase inhibitor、ATPIF1等5个蛋白可作为诱导成骨分化机制研究的候选靶标.
목적 응용iTRAQ기술관찰BMP-2유도기원C2C12세포향성골세포분화과정중단백질표체조적변화.방법 장기원C2C12세포접충우BMP-2유도분화체계중진행분화유도,제취제7천적분화단백이iTRAQ시제표기후진행질보검측,분석차이표체적단백질,병진행생물신식학분석.결과 iTRAQ시제표기적BMP-2유도기원C2C12분화세포단백질표체보분석사선출명현차이표체단백질점위23개,표체상조적단백질점8개,하조적단백질점15개.통과추세분류발현상술차이단백재C2C12세포성골분화적각시기(제1지7천)균존재단백적차이표체.기중부분상조단백재분화조기표체수평승고、부분상조단백재분화만기표체수평승고;동양,부분하조단백재분화조기표체수평하강,부분하조단백재분화만기표체수평하강.초보감정SERCA3、세포색소b5、S100A4、ATPase inhibitor、ATPIF1등5개단백표체출현동태변화,증실상술단백가능여유도성골분화궤제유관.결론 본연구차이단백표체추세적결과현시전정감측C2C12세포성골분화적필요성,제시iTRAQ기술시연구세포분자단백개변적유효적단백질조학방법,SERCA3、세포색소b5、S100A4、ATPase inhibitor、ATPIF1등5개단백가작위유도성골분화궤제연구적후선파표.
Objective To apply iTRAQ technology to observe changes in protein expression group in the process of inducing C2C12 cells differentiation towards osteoblast by BMP-2.Methods The myoblast C2C12 cells were seeded in BMP-2 induced differentiation system for differentiation induction.In the 7th day,differentiation protein was extracted and labeled with iTRAQ reagent.Then,mass spectrometric detection,data analysis of differentially expressed proteins,and analysis of biological information were carried out.Results 23 significantly differentially expressed protein spots were screened by BMP-2-induced myoblast C2C12 differentiated cell protein expression profile analysis,where the protein was labeled with iTRAQ reagent.8 protein points were up-regulated,and 15 protein points were down-regulated.Trend classification found that the above differential protein had differential expression in each period of C2C12 cell osteogenic differentiation (1-7 days).Part of up-regulated protein in the early differentiation period showed high expression level;part of up-regulated protein in the late differentiation period showed high expression level;similarly,part of down-regulated protein in the early differentiation period presented low expression level;part of down-regulated protein in the late differentiation period showed low expression level.Preliminary identification showed SERCA3,Cytochrome bS,S100A4,ATPase inhibitor and ATPIF1 presented dynamic changes,which suggests that these proteins may be related to inducing osteogenic differentiation mechanism.Conclusion The results of differential protein expression trend show the necessity of full monitoring of C2C 12 cells osteogenic differentiation and indicate that iTRAQ technology is an effective method of studying protein changes of cellular molecule.Five proteins including SERCA3,Cytochrome b5,S100A4,ATPase inhibitor and ATPIF1 can be used as candidate targets for osteogenic differentiation mechanism research.