对2株来源于同一亲本细胞但淋巴道转移力显著不同的小鼠肝癌腹水型细胞株Hca-F(淋巴结转移率75%)和Hca-P(淋巴结转移率25%),采用荧光差异双向凝胶电泳(2DDIGE)和DeCyder定量分析软件及HPLC-nESI-MS/MS技术,定量分析和鉴定了小鼠肝癌细胞Hca-F和Hca-P的差异表达蛋白.结果显示,有116个蛋白质点表达水平存在明显差异(P〈0.05),在Hca-F中表达上调蛋白质点62个,下调蛋白质点54个.对所有116个蛋白质点进行了电喷雾串联质谱鉴定,共鉴定出109种单一(Unique)蛋白.其中部分蛋白已被报道与不同类型肿瘤的发生、浸润和转移相关,多数蛋白质被首次报道与肝癌的淋巴道转移过程直接相关.
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 fluo- rescent two-dimensional difference gel electrophoresis (2D DIGE), the expression levels of differentially ex- pressed 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 pro- teins identified in current were for the first time revealed to be involved in the mouse hepatocarcinoma due to the lymphatic metastasis.