肿瘤转移相关基因1在食管癌细胞上皮间质转化过程中的促进作用

The effect of tumor metastasis associated gene 1 on the process of epithelial-mesenchymal transition in esophageal squamous cell carcinoma

目的 探讨肿瘤转移相关基因1(MTA1)对食管癌细胞上皮间质化(EMT)过程的影响及分子机制.方法 采用慢病毒感染方法建立MTA1敲降的食管癌细胞系LV3-shMTA1-KYSE410及MTA1过表达的食管癌细胞系LV5-MTA1-KYSE450;通过Western Blot方法检测上述食管癌细胞系中MTA1蛋白及EMT相关蛋白的表达水平;进而采用免疫荧光实验检测上述食管癌细胞系中EMT相关蛋白分子标志物E-cadherin和Vimentin的定位以及表达水平的变化;最后应用划痕实验明确MTA1表达水平改变后对食管癌细胞迁移能力的影响.结果 食管癌KYSE450细胞系过表达MTA1后,EMT相关蛋白标志物中的E-cadherin表达水平降低而Vimentin表达水平升高,迁移能力增强,细胞迁移率为0.91±0.00,对照组为0.23±0.04;食管癌KYSE410细胞系敲降MTA1后,E-cadherin表达水平升高而Vimentin 表达水平降低,迁移能力降低,细胞迁移率为0.19±0.01,对照组为0.53±0.01,差异均有统计学意义(P均＜0.05).结论 MTA1过表达可促进食管癌细胞的间质化、提高食管癌细胞的迁移能力.

Objective To explore the effect and mechanism of metastasis associated gene 1 (MTA1) on epithelial-mesenchymal transition(EMT) of esophageal squamous cell carcinoma(ESCC).Methods Lentivirus infection method was used to establish the MTA1 knocking out cell line (LV3-shMTA1-KYSE410) and the MTA1 overexpressing cell line (LVS-MTA1-KYSE450).Western Blot was used to measure the expression of MTA1 and the proteins associated with EMT process.Furthermore,the expression and localization of E-cadherin and Vimentin were observed by immunofluorescence assay under confocal microscope.Finally,the wound healing method was performed to confirm the changes of migration ability of the established cell lines.Results When KYSE-450 cells were overexpressed MTA1,the expression level of E-cadherin was down-regulated while Vimentin was up-regulated,and the migration ability was enhanced (0.91 ± 0.00 vs.0.23 ± 0.04,P ＜0.05).When MTA1 was knocked out in KYSE-410 cells,the results were the opposite (0.19±0.01 vs 0.53±0.01,P ＜0.05).Conclusion Overexpression of MTA1 may promote the process of epithelial-mesenchymal transition and enhance the migration ability of ESCC.