目的 探讨Ebp1对人腺样囊性癌(adenoid cystic carcinoma,ACC)细胞迁移的影响及其分子机制.方法 构建Ebp1表达载体pcDNA3.1-Ebp1,建立Ebp1稳定表达细胞系ACC-Ebp1作为Ebp1组,同时设对照组和空载体组.采用细胞体外运动实验系统观察Ebp1对ACC细胞体外运动能力的影响.采用Western blot检测Ebp1过表达后基质金属蛋白酶-9(matrix metalloprotease9,MMP-9)及细胞间黏附分子-1(intercelluar adhesionmolecule 1,ICAM-1)、B钙黏附素(E-cadherin)蛋白表达的变化.结果 侵袭实验中,对照组、空载体组和Ebp1组的穿膜细胞数分别为33±7、92±11和106±5(P=0.03)；运动实验中,3组的穿膜细胞数分别为68±5、168±2和203±4(P=0.02).划痕实验中,对照组与空载体组细胞在第16小时已将划痕填满,而Ebp1组细胞直到第24小时仍未完全填满划痕(P=0.002).与对照组及空载体组相比,Ebp1组ACC细胞中MMP-9的表达降低,而ICAM-1及E-cadherin的表达则增高(P＜0.01).结论 Ebp1过表达有效抑制了ACC细胞的迁移,其作用机制可能与抑制ACC细胞的基质降解能力及增强肿瘤细胞间的黏附有关.%Objective To investigate the effect of Ebpl overexpression on the invasion and motility of human adenoid cystic carcinoma (ACC) cells and its underlying mechanism. Methods Expression vector pcDNA3.1-Ebpl was constructed, and cell lines stably expressed Ebpl was established. Wound health assay and Transwell method were used to examine the invasion and motility of ACC cells. Matrix metalloprotease 9CMMP-9),intercelluar adhesion molecule 1 (ICAM-1) and E-cadherin were detected by Western blot after Ebpl overexpression. Results Numbers of cross-membrane cells in control,vector and Ebpl groups were 33 ± 7,92 ± 11 and 106 ±5 (P = 0.03) in the invasion assay, and 68 ±5,168 ± 2 and 203 ±4 (16 fields,P = 0.02) in the motility assay. Control and vector groups both took 16 hours for wound health, while the time in Ebpl group was more than 24 hours (P = 0. 002). After Ebpl overexpression,MMP-9 was downregulated, while ICAM-1 and E-cadherin were upregulated (P< 0.01). Conclusions Ebpl inhibits the migration ability of ACC cells, which may be associated with decreasing of matrix degradation, enhancement of adhesion among cells and inhibition of epithelial mesenchymal transition.
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