摘要目的 研究烯酯酰辅酶A水解酶1( ECH1)在小鼠肝癌高、低淋巴转移Hca-F/Hca-P细胞株中的表达及ECH1基因表达下调后对Hca-F细胞增殖和迁移能力的影响.方法 采用免疫荧光方法检测ECH1在小鼠肝癌细胞株中的表达.构建pGPU6/GFP/Neo-shRNA-ECH1表达载体,稳定转染至高淋巴转移Hca-F细胞中干扰ECH1基因的表达,通过CCK8法检测肿瘤细胞的增殖能力,Transwell小室检测肿瘤细胞的迁移能力,并应用流式细胞仪检测肿瘤细胞的细胞周期情况.结果 ECH1在小鼠肝癌细胞株中主要表达于细胞质,且ECH1在Hca-F细胞的表达强于其在Hca-P细胞的表达.ECH1表达下调后,Hca-F细胞的增殖能力明显下降.Transwell小室检测穿膜细胞数pGPU6/GFP/Neo-shRNA-ECH1转染Hca-F组(27.07±17.49)较阴性对照组(72.38±18.83)和Hca-F组(59.06±30.33)明显减少,细胞周期S期pGPU6/GFP/Neo-shRNA-ECH1转染Hca-F组(86.1％)较阴性对照组(75.8％)和Hca-F组(66.2％)停留细胞数明显增多,G1期pGPU6/GFP/Neo-shRNA-ECH1转染Hca-F组(9.4％)较阴性对照组(24.2％)和Hca-F组(30.3％)停留细胞数明显减少.结论 ECH1可能在肿瘤淋巴转移中发挥作用,抑制ECH1的表达可有效降低体外高淋巴转移肿瘤细胞的增殖和迁移能力.

abstractsObjective To study the expression of enoyl CoA hydratase 1 ( ECH1 ) and the effect when down-regulation of ECH1 gene expression in mouse hepatocarcinoma cell. Methods 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.Results 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 ),decease 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 G1 phase (9.4％) compared with scramble-negative (24.2％ ) and Hca-F controls (30.3％ ).Conclusion ECH1 serves as a potential critical factor attributes to tumor lymphatic metastasis.