摘要目的 研究细胞内氯离子通道蛋白1(CLIC1)在小鼠腹水型肝癌高、低淋巴道转移细胞株中的定位及表达差异.方法 以小鼠腹水型肝癌高淋巴道转移细胞株Hca-F和低淋巴道转移细胞株Hca-P为研究对象,采用双向凝胶电泳和质谱鉴定、细胞免疫荧光、细胞免疫化学染色方法和Western blot方法检测CLIC1在高、低淋巴道转移细胞株中的定位及蛋白表达情况.结果 双向凝胶电泳和质谱鉴定CLIC1在Hca-F细胞中高表达,是Hca-P细胞的1.6倍;细胞免疫荧光和免疫化学结果显示CLIC1定位于Hca-F和Hca-P细胞的细胞质和细胞膜,功能形式的细胞膜定位更多存在于Hca-F细胞中,并且在Hca-F细胞中的表达强于Hca-P细胞.Western blot结果显示CLIC1在Hca-F细胞中的表达是Hca-P细胞的1.6倍.结论 CLIC1在Hca-F和Hca-P细胞中定位于细胞质和细胞膜,在Hca-F细胞中高表达,且在Hca-F中更多定位于细胞膜,可能在肝癌淋巴道转移过程中发挥作用.

abstractsObjective To study the localization and expression of CLIC1 in mouse hepatocarcinoma ascites cell lines with different metastatic potentials. Methods Mouse heptocarcinoma ascites models (a high potential of lymphatic metastasis cell line-Hca-F,and a low potential of lymphatic metastatsis cell line-Hca-P)were investigated using fluorescent two-dimensional difference-gel electrophoresis (2-D DIGE)and mass spectrometry for detecting the localization and expression of CLIC1. Immunofluore-scence,immunocytochemistry and Western blot were used to assess CLIC1 protein status in the two cell lines.Results CLIC1 expression was obtained in the cytoplasm and plasma membrane of cells in both cell lines. 2-D DIGE showed that CLIC1 was overexpressed in Hca-F cells,1. 6 folds higher than that of the Hca-P cells. Hca-F cells also had a higher integral membrane CLIC1 in the Hca-P cells. Conclusions Although CLIC1 expression is detected in both Hca-F and Hca-P cell lines,a higher protein expression level is present in Hca-F cells. CLIC1 may play an important role in tumor metastasis.