Abstract: Two mouse hepatocarcinoma ascites syngeneic cell lines, a Hca-F with lymph node metastasis rate of 75% and a Hca-P with low lymph node metastasis rate of 25%were established and well maintained in our laboratory. The different expressed proteins between the two cell lines were separated and compared with fluorescent two-dimensional difference gel electrophoresis(2D DIGE), the expression levels of differentially expressed proteins were quantified by DeCyder software and protein identifications of interested protein spots were performed via the high performance liquid chromatography-nano electrospray HPLC-nESI-MS/MS approach. Among the total 116 protein spots obtained by the 2D DIGE, 62 protein spots were observed up-regulated in Hca-F and 54 protein spots up-regulated in Hca-P cell lines, respectively. 109 unique proteins were identified from all the 116 different expressed protein spots. Part of the identified protein candidates have been reported to be associated with the occurrence, invasion and metastasis of a variety of tumors. However, most of the proteins identified in current were for the first time revealed to be involved in the mouse hepatocarcinoma due to the lymphatic metastasis.

Key words: Fluorescent two-dimensional difference gel electrophoresis, HPLC-nESI-MS/MS, Hepatocarcinoma, Lymph node metastasis