探究转录因子FOXM1不同的剪接异构体对乳腺癌EMT过程中的影响.采用基因工程方法分别构建了表达FOXM1—EGFP和FOXM1C—EGFP两种FOXM1剪接异构体真核表达质粒,并将其转染进乳腺癌细胞,采用RT—PCR和Western印迹检测细胞样本中FOXM1剪接异构体的表达和EMT相关基因表达,同时采用transwell检测高表达不同FOXM1剪接异构体细胞的侵袭和迁移能力.成功构建了FOXM1B-EGFP和FOXM1C—EGFP真核表达质粒.外源FOXM1在乳腺癌间质型细胞的表达高于上皮型细胞,并主要存在于细胞核内,且高表达FOXM1B能够显著促进乳腺癌细胞的侵袭和EMT过程.外源FOXM1C在乳腺癌上皮型细胞中的表达高于间质型细胞,并且在细胞核和细胞质中均有表达,高表达FOXM1C能够显著抑制细胞的侵袭和EMT过程.实验结果表明,在乳腺癌细胞中,FOXM1B主要存在于细胞核内,FOXM1C在细胞核和细胞质中均有表达.本研究预示FOXM1B和FOXM1C对乳腺癌细胞EMT过程发挥不同的影响.
To explore the impact of different transcription factor FOXM1 isoforms in breast cancer EMT process, the eukaryotic expression plasmids for two FOXM1 isoforms, FOXM1B-EGFP and FOXMIC-EGFP, were constructed and transfected into breast cancer cells. The expression levels of FOXM1 isoforms and EMT related genes in the cells were detected with RT-PCR and Western blot. The migration ability of the cells overexpressing the FOXM1 isoforms was measured with the transwell test. The FOXMIB-EGFP and FOXMIC-EGFP eukaryotic expression plasmids were successfully constructed. We found that the levels of exogenous FOXM1B in mesenehymal cells were higher than those in epithelial cells, and it was mainly located in the nucleus. The high levels of FOXM1B expression significantly stimulated the invasion of breast cancer cells and EMT process. The levels of exogenous FOXM1C in epithelial cells were higher than those in mesenchymal cells, and they were expressed in both the nucleus and the cytoplasm. The high levels of FOXMIC expression inhibited the invasion of breast cancer cells and EMT process. FOXM1B was located mainly in the nucleus of cells and FOXMIC was expressed in both the nucleus and the cytoplasm of cells. The research has indicated that FOXMIB and FOXM1C play different roles in the process of EMT of breast cancer cells.