Background and purpose: Our previous study have identified many lymphatic metastais-assoeiated gencs/protcins using gene chip and quantitative protconiics technology from a couple of the same parents' mouse hepatocarcinoma cell lines with high and low lymphatic metastalic potential. In this study, we select chloride intracellular channel 1 (CLJC1) which is highly expressed in cells with high metastatie potential to understand the effect of CLIC1 overexpression on tumor biological behavior in mouse hepatocarcinoma cell line Hca-P in vitro. Methods: The plasmids pcDNA3.1(+)-CLICl was constructed. Then the Hca-P cells were transfected with the pcDNA3.1(+) and pcDNA3.1(+)-CLIC1. The stably transfected cells were obtained by growing them in G418 for 4 weeks. The levels of expression of CLIC1 mllNA and protein were detected by semi-quantitative polymerase chain reaction ('RT-PCR) and enzyme-linked immimosorbent assay (ELISA) analysis, respectively. The cell proliferation was detected by CCK-8 method; the invasive and migrative abilities were detected by Transwell assay. Results: Restrictive endonuclease assay and sequence analysis verified the successful construction of the recombinant vector pcDNA3.1 (+)-CLlCl. The cell line Hca-P that steadily expressing CLIC1 gene was constructed. RT-PCR assay and ELISA showed that the expression level of CLICI mRNA and protein in the I Ica-P cells were increased significantly after pcDNA3.1(i )-CLICI transfection (P<0.05). The cell proliferation, the average number of invasive and migrative cells were increased in Hca-P cells with CLICI transfection as compared with that of the Hca-P without CLICI transfection (P展开▼
