ABSTRACT
<p><b>OBJECTIVE</b>To study the expression of enoyl CoA hydratase 1 (ECH1) and the effect when down-regulation of ECH1 gene expression in mouse hepatocarcinoma cell.</p><p><b>METHODS</b>Immunofluorescence was used for detecting the expression of ECH1, and stably transfected Hca-F cells with pGPU6/GFP/Neo-shRNA-ECH1 expression plasmids. Cell proliferation was assessed by Cell counting kit-8 (CCK8) assay. The Boyden-transwell assay (8 µm pore size) was performed to analyze the inhibitory effect of shRNA on Hca-F cell migration and invasion.</p><p><b>RESULTS</b>ECH1 expression was obtained in the cytoplasm and upregulated expression in Hca-F cells than that in Hca-P cells. The down-regulation of ECH1 could inhibit the cell proliferation of Hca-F cells, decrease the number of cell pass through Transwell (27.07 ± 17.49) compared with scramble-negative (72.38 ± 18.83) and Hca-F controls (59.06 ± 30.33), decrease the migration capacities of Hca-F cells, increase the ratio of Hca-F cells in S phase (86.1%) compared with scramble-negative (75.8%) and Hca-F controls (66.2%) and decrease the ratio of G(1) phase (9.4%) compared with scramble-negative (24.2%) and Hca-F controls (30.3%).</p><p><b>CONCLUSION</b>ECH1 serves as a potential critical factor attributes to tumor lymphatic metastasis.</p>
Subject(s)
Animals , Mice , Cell Cycle , Cell Line, Tumor , Cell Movement , Cell Proliferation , Cytoplasm , Down-Regulation , Enoyl-CoA Hydratase , Genetics , Metabolism , Gene Expression Regulation, Neoplastic , Liver Neoplasms, Experimental , Pathology , Lymphatic Metastasis , Plasmids , RNA, Small Interfering , Genetics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To study the effects of silencing CLIC1 gene expression on the proliferation and invasion of Hca-F cells.</p><p><b>METHODS</b>The mouse CLIC1 cDNA sequence was retrieved from NCBI. Three shRNA sequences were designed and cloned into pGPU6/GFP/Neo plasmids. The plasmids were transfected into Hca-F cells with Lipofectamine 2000. Cell Counting-8 (CCK-8) kit and transwell chamber were used to study the effects of CLIC1 on the proliferation and invasion of Hca-F cells.</p><p><b>RESULTS</b>The pGPU6/GFP/Neo-shRNA-3 plasmid effectively repressed the expression of CLIC1 mRNA. Inhibition of CLIC1 gene expression led to decreased cell proliferation and reduced invasion.</p><p><b>CONCLUSION</b>CLIC1 is essential for the proliferation and invasion of Hca-F cells.</p>