摘要
目的:筛选与鉴定胰腺癌干细胞相关的差异表达蛋白质.方法:以MIA-PaCa2(TIChigh)与BxPc-3(TIClow)为研究胰腺癌干细胞的工具细胞,采用荧光差异双向凝胶电泳技术分离并筛选上述细胞差异表达蛋白,反射式基质辅助激光解吸附电离串联飞行时间质谱鉴定差异蛋白,免疫印迹验证差异表达的TRIM28.结果:获得了MIA-PaCa2(TIChigh)与BxPc-3(TIClow)荧光差异蛋白表达图谱,经DeCyder v6.5软件分析,共有23个差异在1.5倍以上的蛋白质点,经质谱鉴定得到19个蛋白质,相对于BxPc-3(TIClow),在MIA-PaCa2(TIChigh)细胞组中高表达者有8个,低表达者有11个,包括参与细胞通讯和信号传导蛋白、免疫反应蛋白、转录因子与细胞周期调控蛋白、调节脂肪细胞分化和脂滴形成蛋白、细胞骨架蛋白、细胞黏附分子、差向异构酶、转运活性蛋白及参与翻译调节相关蛋白.免疫印迹结果显示TRIM28在BxPc-3(TIClow)没有表达,在MIA-PaCa2(TIChigh)高表达.结论:筛选得到的TRIM28等胰腺癌干细胞相关差异表达蛋白可能与胰腺癌干细胞的发生、发展和调控机制相关,有望成为胰腺癌新的治疗靶点.
AIM： To screen and identify differentially expressed proteins in pancreatic cancer stem cells. METHODS： MIA-PaCa2 （TIChigh） and BxPc-3 （TIClow） were used in the study. Differentially ex pressed proteins between MIA-PaCa2 （TICghih） and BxPc-3 （TIClow） cells were isolated and screened by 2D-DIGE analysis. Protein identification was performed by peptide mass fingerprinting with matrix-assisted laser desorption/ionization time- of-flight mass spectrometry （MALDI-TOF/TOF）. Western blot was performed to verify the differential expression of TRIM28.RESULTS： Fluorescent differential protein expression patterns were obtained between MIA-PaCa2 （TIChigh） and BxPc-3 （TIClow） cells. Analyses with DeCyder v6.5 software showed a total of 23 differentially expressed protein spots （〉1.5 folds）, and these protein spots were identified by mass spectrometry as 19 proteins, which are involved in cell communication and signal transduction, immune response, transcription and cell cycle regulation, adipocyte differentiation and lipid droplet formation, cytoskeletal formation, cell adhesion, transport, and translation. Western blot analysis revealed that TRIM28 was highly expressed in MIA-Pa- Ca2 （TIChigh） cells but not expressed in BxPc-3 （TIClow） cells. Among the 19 identified proteins, 8 were up-regulated and 11 down-regulated in MIA-PaCa2 （TIChigh） cells.CONCLUSION： The identified differentially expressed proteins, such as TRIM28, are associated with the genesis, development and regulation of pancreatic cancer stem cells. They may become new therapeutic targets for pancreatic cancer.
出处
《世界华人消化杂志》
CAS
北大核心
2013年第2期145-152,共8页
World Chinese Journal of Digestology
基金
国家自然科学基金资助项目
No.81160311
贵州省科技厅贵阳医学院社发联合基金资助项目
No.黔科合[2010]3171
贵阳市科技局基金资助项目
No.筑科合[2011103]22号
贵州省肝胰疾病研究科技创新人才团队基金资助项目
No.黔科合人才团队[2010]4010~~
关键词
胰腺癌
干细胞
电泳
凝胶
双向
蛋白质组学
Pancreatic carcinoma
Stem cells
Electrophoresis
Gel
Two-dimensional
Proteomics
