紫杉醇抗胆管癌细胞差异表达蛋白的分离、质谱鉴定及生物信息学分析

目的：筛选紫杉醇诱导胆管癌细胞凋亡的相关蛋白并鉴定差异表达的蛋白点以寻找抗胆管癌药物作用的新靶点，筛选胆管癌新的肿瘤标志物，为胆管癌的早期诊断、疗效观察提供基础理论依据。方法：应用比较蛋白质组学技术对紫杉醇诱导凋亡的胆管癌QBC939细胞进行蛋白分离和蛋白表达谱差异分析，筛选并鉴定其相关蛋白并测定其胶内酶解后的肽指纹图谱。结果：本研究发现对照组和实验组蛋白点匹配率分别为（90．1±0．14）％和（87．1±0．22）％（P〈0．05），匹配后在干预组中共发现68个差异表达蛋白点，其中47个表达下调和21个上调；共获得28张PMF，初步质谱鉴定发现11个与凋亡相关的差异表达蛋白，其中6种蛋白表达上调，5种蛋白表达下调。结论：应用蛋白质组学技术分析所获得的68个差异蛋白质点可能在QBC939细胞发生凋亡过程中发挥重要作用；被鉴定出的11个与细胞凋亡相关的差异蛋白点提示紫杉醇是通过多个重要蛋白，发挥其诱导细胞凋亡的抗癌作用机制的。

To identify and separate differential expression proteins in cholangiocarcinoma cells treated with paclitaxel by mass spectra and bioinformatics

Objective：To investigate new early tumor diagnosis markers for cholangiocarcinoma and new molecule mechanism of inducing with paclitaxel and new therapy target in curing cholangiocarcinoma. To provide theory foundation for therapeutic effect observation by identification of the differential expression protein spots. Methods： Introduce two dimensional gel electrophoresis and computer - assisted image analysis to seperate and assess the apoptosis - associated proteins in QBC939 treated by paclitaxel. To detect peptide mass fingerprinting with MALDI - TOF - MS after spots enzymolyed. Results：The matching ratio of the protein spots in experiment group and the control group were （90.1 ± 0.14 ） % and （ 87.1 ± 0.22 ） % respectively （ P 〈 0.05 ）. There were 68 protein spots at different level of expression,including 47 increased and 21 decreased in interfered groups. Twenty - eight pieces of PMF were matched , among the identified protein spots the expression level of protein spot was up - regulated including 6 proteins and downregulated including 5 proteins. Conclusion：Total 68 different expression of protein spots may play an important role in QBC939 cell lines induced apoptosis by paclitaxel. Total 11 protein spots related to apoptosis may involve in the anticancer mechanism of induced apoptosis.