胃癌不同转移能力细胞系的2-DE图谱及差异分析

2-DE profiling and differential analysis of highly and poorly metastatic human gastric carcinoma cell lines

目的建立胃癌低转移能力细胞系RF1和高转移能力细胞系RF48的2DE图谱,并分离胃癌转移相关蛋白质。方法利用固相pH梯度技术(immobilizedpHgradient,IPG)双向凝胶电泳(twodimensioanlpolyacrylamidegelelectrophoresis,2DE)建立胃癌低转移能力细胞系RF1和高转移能力细胞系RF48二维电泳图谱;采用图像分析软件进行比较分析,识别差异表达的蛋白质,分离胃癌转移相关蛋白质。再利用基质辅助激光解析电离飞行时间质谱(MALDITOFMS)及数据库查询,对部分差异表达蛋白质点进行鉴定。结果建立胃癌低转移能力细胞系RF1和胃癌高转移能力细胞系RF48双向凝胶电泳图谱;三块凝胶的平均蛋白质点数分别为960±92和978±83,平均匹配的点数分别为780±69和769±53,匹配率达87%和86.3%;对25个差异蛋白质点进行肽质指纹图分析,鉴定出与瘤基因、细胞周期调控和信号转导有关的蛋白质。结论建立分辨率高且重复性较好的人胃癌低转移能力细胞系RF1和胃癌高转移能力细胞系RF48的双向凝胶电泳图谱,可以识别和鉴定出一些与胃癌转移相关的差异表达蛋白质。

Purpose To establish two-dimensional electrophoresis profiles with high resolution and reproducibility from RF-1 cell line （primary tumor of a gastric adenocarcinoma patient） and RF-48 cell line its metastatic counterpart, and to identify gastric adenocarcinoma metastasis related proteins. Methods Immobilized pH gradient-two dimensional polyacrylamide gel electrophoresis, silver staining, PDQuest 2-DE software analysis, peptide mass fingerprinting based on matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS） and SWISS-PROT database searching, were used to separate and identify the differential proteomic expressions between RF-1 cell line and RF-48 cell line. Results The good. 2-DE pattern between RF-1 cell line and RF-48 cell line, including high resolution and reproducibility was obtained. The average Protein spots of 3 gels were 960 ± 92, and 978± 83. The average matching spots were 780 ±69, and 769 ±53. Spots were matched with the average matching rate of 87% and 86. 3%. 25 differential spots （ 16 spots in RF-1 and 9 spots in RF-48 ） were analysed with Pept Ident software. 8 protein were preliminarily identified. These proteins were related to cell signal transduction, cell metabolism, proliferation and differentiation etc. Conclusions The wellresolved, reproducible 2-DE patterns of RF-1 cell line and RF-48 cell line were established and certain differential proteins are characterized. Some proteins relate to metastasis are preliminarily scanned.