Tanaka Toshiyuki, Kuramitsu Yasuhiro, Fujimoto Masanori, Naito Seiji, Oka Masaaki, Nakamura Kazuyuki
Department of Digestive Surgery and Surgical Oncology Department of Surgery, Yamaguchi University Graduate School of Medicine, Yamaguchi, Japan.
Electrophoresis. 2008 Jun;29(12):2651-9. doi: 10.1002/elps.200700847.
Proteomic differential display analysis was performed on human renal cell carcinoma cell SN12C clones having different metastatic potentials by using 2-DE and LC-MS/MS. The SN12C cell clones were SN12C parent cell line, SN12C-clone 2, SN12C-clone 4, and SN12C-PM6. The SN12C parent cell line was established from an HRCC surgical specimen. SN12C-clone 4 has lower, and SN12C-clone 2 and SN12C-PM6 have higher metastatic potential than SN12C parent cells. We found eight protein spots whose expression level was different between low metastatic clones and high metastatic clones. The protein expression of three appeared to be higher in high metastatic clones than low metastatic clones, and that of other five protein spots appeared to be lower in high metastatic clones than low metastatic clones. These spots were selected, digested and analyzed by LC-MS/MS analysis, and they were identified by peptide sequencing tag. In high metastatic potential clones, two isoforms of ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCH-L1) were downregulated. These results suggest that UCH-L1 expression seems to be associated with the metastatic potential of HRCC SN12C cell clones.
通过二维电泳（2-DE）和液相色谱-串联质谱（LC-MS/MS）技术，对具有不同转移潜能的人肾癌细胞SN12C克隆进行了蛋白质组差异显示分析。SN12C细胞克隆包括SN12C亲本细胞系、SN12C-克隆2、SN12C-克隆4和SN12C-PM6。SN12C亲本细胞系源自一份肾透明细胞癌（HRCC）手术标本。与SN12C亲本细胞相比，SN12C-克隆4的转移潜能较低，而SN12C-克隆2和SN12C-PM6的转移潜能较高。我们发现了8个蛋白质点，其在低转移克隆和高转移克隆之间的表达水平存在差异。其中3个蛋白质点在高转移克隆中的表达似乎高于低转移克隆，另外5个蛋白质点在高转移克隆中的表达似乎低于低转移克隆。选取这些蛋白质点进行消化，并通过LC-MS/MS分析，通过肽序列标签进行鉴定。在高转移潜能克隆中，泛素羧基末端水解酶同工酶L1（UCH-L1）的两种同工型表达下调。这些结果表明，UCH-L1的表达似乎与HRCC SN12C细胞克隆的转移潜能相关。
