摘要
通过比较研究不同转移潜能肝癌细胞系中核心岩藻糖基化蛋白质表达谱的差别,筛查与转移相关的重要糖蛋白.用SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)、双向电泳(2-DE)和凝集素印迹技术联合基质辅助激光解吸飞行时间串联质谱(MALDI-TOF-MS/MS)分析,建立3种不同转移潜能人肝癌细胞系Hep3B、MHCC97L和MHCC97H的核心岩藻糖基化蛋白质表达图谱.比较研究发现,不同转移潜能肝癌细胞呈现不同的SDS-PAGE/LCA凝集素印迹图谱,MHCC-97H和MHCC-97L在35～45 ku和45～60 ku间出现了Hep3B未见的条带.在核心岩藻糖基化蛋白质表达图谱中,Hep3B、MHCC97L和MHCC97H分别平均检测到(55±7)个蛋白质点(n=3),(60±6)个蛋白质点(n=3),(61±4)个蛋白质点(n=3);以各自双向电泳图谱为参考胶,Hep3B、MHCC97L和MHCC97H分别与其匹配的平均匹配点数为(25±3)个(n=3),(30±4)个(n=3),(28±3)个(n=3).该图谱中,与Hep3B相比,MHCC97L有13个点未匹配,其中9个点为Hep3B(-)/MHCC97L(+);MHCC97H有9个点未匹配,其中6个点为Hep3B(-)/MHCC97H(+),MALDI-TOF-MS/MS可鉴定出Annexin1、Keratin 8等12种差异蛋白质.这些结果证实了不同转移潜能的肝癌细胞有明显的核心岩藻基化糖蛋白差异性表达.提示肝癌转移可能与这些差异糖蛋白及其核心岩藻糖基化有关.
In order to comparatively analyze the core-fucosylated glycol-proteome expression profile of hepatocellular carcinoma cell lines (HCC) with different metastasis potentials, some key glycoprotein correlated with the metastasis of hepatoma were screened. Using SDS-PAGE, 2-DE followed by LCA lectin blot, differential display maps of core-fucosylated glycol-proteome were generated and analyzed, and then the glycoproteins were identified by a matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS/MS). I-DE lectin blot showed that MHCC97-H and MHCC97-L, which were established by authors' institute and had higher metastasis potentials than Hep3B, displayed numerous bands. From the core-fucosylated glycol-proteomics expression profiles, (55 +/- 7) spots in Hep3B (n=3), (60 +/- 6) spots in MHCC97-L(n=3), and (61 +/- 4) spots in MHCC97-H (n=3) were detected. Matched with individual 2-DE maps, the core-fucosylated glycol-proteomics expression profiles showed that there were (25 +/- 3) detected spots in Hcp3B (n=3), (30 +/- 4) detected spots in MHCC97-L and (28 +/- 3) detected spots in MHCC-97H (n=3). There were 13 unmatched spots between the expression profile of Hep3B and NMCC97-L, and 9 unmatched spots between Hep3B and MHCC97-H. The 12 differential glycoproteins were identified by MALDl-TOF-MS/MS. Individually differential core-fucosylated glycoproteomics expression profiles existed in HCC with different metastasis potentials. All the results suggested that the metastasis of HCC might be correlated with these differential glycoproteins.
出处
《生物化学与生物物理进展》
SCIE
CAS
CSCD
北大核心
2005年第5期442-448,共7页
Progress In Biochemistry and Biophysics
基金
国家高技术"863"计划资助项目(2002BA711A11)
国家重点基础研究发展规划项目(973)(2001CB510205
2004CB520802)~~
