细胞内氯离子通道蛋白1在肝癌高、低淋巴道转移细胞株中的定位及表达差异

Localization and expression of CLIC1 in hepatocarcinoma ascites cell lines with high or low potentials of lymphatic spread

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Westernblot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CLICl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CLICl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。Westernblot结果显示CLICl在Hca—F细胞中的表达是Hca—P细胞的1．6倍。结论CLICl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。

Objective 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, immunocytoehemistry 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 ceils 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.