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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MDAMB-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 analyze the role of Snail and MTDH genes in breast cancer cells and determine whether Snail promotes breast cancer metastasis by binding to the promoter region of MTDH. Methods The Snail gene was transfected into breast cancer cells and the changes of MTDH mRNA and protein expression in Snail-overexpressing breast cancer cells were observed. Co-immunoprecipitation was used to detect the co-effect of Snail and MTDH genes. Results After transfection of Snail into breast cancer cell line MDAMB-435, the MTDH mRNA expression levels in transfection group, blank group and control group were 1.61 ± 0.22, 1.02 ± 0.18 and 0.99 ± 0.20 respectively, (P <0.05), while there was no significant difference between the two groups (P> 0.05). Western blot results showed that Snail promoted the expression of MTDH protein. Co-immunoprecipitation showed that Snail and MTDH in the cells Memory is combined with each other. Conclusion Snail can promote the metastasis of breast cancer by binding to the promoter region of MTDH gene in breast cancer cells, promoting the transcription of MTDH mRNA and the expression of related proteins.