靶向下调FOXM1 基因表达对乳腺癌MDA-MB-231细胞增殖的影响

目的：探讨靶向抑制FOXM1 对乳腺癌细胞增殖能力的影响，为乳腺癌的个性化靶向治疗提供理论依据。方法：利用重组真核转录载体pSilencer1.0-U6-FOXM1-shRNA，脂质体法转染乳腺癌细胞株MDA-MB-231，下调其FOXM1基因表达。采用四甲基偶氮唑盐(MTT) 比色法、平板克隆形成实验观察细胞增值曲线以及克隆形成能力；采用实时定量- 聚合酶链反应(Real-timeqPCR)、蛋白免疫印迹法（Western blot）分别检测FOXM1 基因在mRNA、蛋白水平的表达变化。结果：重组载体pSilencer1.0-U6-FOXM1-shRNA转染MDA-MB-231细胞后，与对照组相比，增殖速率明显下降（P<0.05），平板克隆形成显著减少（P<0.05），重组载体转染后显著抑制MDA-MB-231 细胞中FOXM1 基因在mRNA、蛋白水平的表达。结论：沉默FOXM1 基因对乳腺癌细胞株MDA-MB-231 生长具有抑制作用，为阐明乳腺癌发病机制提供了新的切入点，也为临床抑制肿瘤生长提供了新的作用靶点。

Endothelial Growth Factor

This experiment is designed to figure out the effect of breast cancer cells proliferation after target silencingof FOXM1, thus providing a mineral strategy for individualized and target breast cancer therapy. The recombinant vectors withshRNA targeting FOXM1 (pSilencer1.0-U6-FOXM1-shRNA) were transfected into MDA-MB-231 cells with the help of lippofectamine2000 to down regulate the expression of FOXM1. MTT assay and Colony Forming Assay were used to evaluate the proliferation andcolony forming efficacy of MDA-MB-231 cells. Meantime, Real-Time quantitative Polymerase Chain Reaction (RT-qPCR) and Westernblot assay were performed to monitor the expression of mRNA and protein level of FoxM1. After transfection of the pSilencer1.0-U6-FOXM1-shRNA target FOXM1, cell proliferation was enormously decreased compared to control groups (P<0.05). FOXM1expression in the transfected MDA-MB-231 cells was significantly depressed both at mRNA and protein level. ThepSilencer1.0-U6-FOXM1-shRNA target FOXM1 could efficiently inhibit FOXM1 gene expression in breast cancer cell lineMDA-MB-231. Stable silence of FOXM1 gene by shRNA could negatively regulate cell proliferation in human breast cancerMDA-MB-231 cells. This experiment could provide a new insight into the mechanism of breast cancer and suggest a promising target ofsuppression cancer growth.