肝癌转移相关的核心岩藻糖基化 蛋白质表达谱的研究

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 .

Core-fucosylated Glycoproteomics Correlated With The Metastasis of Hepatoma

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). 1-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 Hep3B(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 MHCC97-L, and 9 unmatched spots between Hep3B and MHCC97-H. The 12 differential glycoproteins were identified by MALDI-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.