目的 分析乳腺癌细胞中Snail与MTDH基因的作用,明确Snail是否通过结合于MTDH的启动子区域促进乳腺癌转移。方法 克隆、转染Snail基因至乳腺癌细胞,观察过表达Snail的乳腺癌细胞中MTDHmRNA及蛋白表达的变化;再使用免疫共沉淀法检测Snail与MTDH基因的共作用。结果 转染Snail基因进入乳腺癌MDA-MB-435细胞后,转染组、空白组和对照组中MTDHmRNA的表达水平分别为1.61±0.22、1.02±0.18、0.99±0.20,转染组高于空白组和对照组,差异有统计学意义(P<0.05),而后两组表达无差异(P>0.05);Westren blot检测结果显示,Snail可促进MTDH蛋白的表达;免疫共沉淀显示,Snail与MTDH在细胞内存在相互结合作用。结论 Snail在乳腺癌细胞中可通过结合于MTDH基因的启动子区域,促进MTDHmRNA转录及相关蛋白的表达,从而导致乳腺癌转移。

Objective To investigate the function of Snail to MTDH gene in breast cancer cells. Methods We observed the changement of MTDHmRNA and protein expression in breast cancer cell line MDA-MB-435 after transfected with Snail gene. Then, we used co-immunoprecipitation to determine the domain of Snail and MTDH binding in vitro. Results After transfected with Snail gene into MDA-MB-435 cell, the expression levels of MTDHmRNA in transfection group, blank group and control group were 1.61±0.22,1.02±0.18,0.99±0.20. The level of transfection group was significantly higher than the other groups(P< 0.05). Western blot showed that the expression of MTDH protein can be promoted by Snail. Co-immunoprecipitation showed that Snail and MTDH are binding interactions in breast cancer cell line MDA-MB-43. Conclusion Snail can promote transcription and expression of MTDH in breast cancer cells by binding to the promoter region of the MTDH gene resulting in metastasis of breast cancer.