烯酯酰辅酶A水解酶1基因在小鼠肝癌细胞株中的表达及其与增殖和迁移能力的关系  

Expression of enoyl CoA hydratase 1 reduces cell proliferation and migration in mouse hepatocarcinoma cells

作　　者：张军[1] 唐建武[1] 宋美英[1] 王波[1] 李荣宽[1] 侯力[1] 黄玉红[1] 王梅[1] 宋波[1] 金艳玲[1] 

机构地区：[1]大连医科大学病理学教研室,116044

出　　处：《中华病理学杂志》2011年第12期830-833,共4页Chinese Journal of Pathology

基　　金：国家自然科学基金（81071725）;辽宁省教育厅高等学校科研项目（2009S028）;辽宁省科学技术计划项目（2008225010-3）

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.Objective 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.

关 键 词：肝肿瘤 烯酰CoA水合酶 显微镜检查 荧光 淋巴转移 

分 类 号：R735.7[医药卫生—肿瘤]