Zhang Jun, Tang Jian-wu, Song Mei-ying, Wang Bo, Li Rong-kuan, Hou Li, Huang Yu-hong, Wang Mei, Song Bo, Jin Yan-Ling
Department of Pathology, Dalian Medical University, Dalian, China.
Zhonghua Bing Li Xue Za Zhi. 2011 Dec;40(12):830-3.
To study the expression of enoyl CoA hydratase 1 (ECH1) and the effect when down-regulation of ECH1 gene expression in mouse hepatocarcinoma cell.
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.
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%).
ECH1 serves as a potential critical factor attributes to tumor lymphatic metastasis.
研究烯酰辅酶A水合酶1（ECH1）的表达以及ECH1基因表达下调对小鼠肝癌细胞的影响。
采用免疫荧光法检测ECH1的表达，并将pGPU6/GFP/Neo-shRNA-ECH1表达质粒稳定转染至Hca-F细胞。通过细胞计数试剂盒-8（CCK8）法评估细胞增殖。采用Boyden迁移小室实验（孔径8μm）分析shRNA对Hca-F细胞迁移和侵袭的抑制作用。
ECH1在细胞质中表达，且在Hca-F细胞中的表达上调，高于Hca-P细胞。ECH1表达下调可抑制Hca-F细胞的增殖，与阴性对照（72.38±18.83）和Hca-F对照（59.06±30.33）相比，穿过Transwell的细胞数量减少（27.07±17.49），降低Hca-F细胞的迁移能力，与阴性对照（75.8%）和Hca-F对照（66.2%）相比，增加Hca-F细胞S期的比例（86.1%），与阴性对照（24.2%）和Hca-F对照（30.3%）相比，降低G(1)期的比例（9.4%）。
ECH1作为一个潜在的关键因素与肿瘤淋巴转移有关。
