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目的:探究转录因子HOXA5在乳腺癌进展中的作用。方法:构建过表达HOXA5的稳定转染BT549和SUM159细胞系,在稳转BT549细胞系中利用Transwell检测细胞的转移能力;Western blotting检测上皮-间质转化(EMT)相关蛋白表达水平的变化;平板克隆实验检测细胞的增殖能力;转录组测序技术(RNA-seq)进一步分析HOXA5的调控基因,利用软件Cluster 3.0,treeview和DAVID生物信息数据库(DAVID Bioinformatics Resources)以及Enrichr分析KEGG信号通路,绘制热图(heatmap)。结果:BT549细胞系中,稳定转染HOXA5后的实验组细胞迁移能力显著降低(P<0.001),且上皮标志物E-cadherin蛋白表达水平显著上调,间质标志物N-cadherin,Twist1,Slug蛋白表达水平显著下调;平板克隆结果表明,实验组形成的克隆数目明显减少(P<0.05),克隆大小明显降低。结论:HOXA5通过促进细胞由间质向上皮转化,抑制乳腺癌细胞迁移,并且抑制细胞增殖;HOXA5通过调节糖脂代谢途径调节癌细胞增殖,并且还通过调节细胞运动和粘附相关的基因抑制肿瘤细胞的迁移,通过肿瘤坏死因子(TNF)信号通路发挥抗肿瘤作用。总之,转录因子HOXA5对乳腺癌的发展和恶化起着显著的抑制作用。
Objective: To investigate the role of transcription factor HOXA5 in the progression of breast cancer. Methods: Transfection of BT549 and SUM159 cell lines stably transfected with HOXA5 was established. Transwell assay was used to detect cell viability in BT549 cells. Western blotting was used to detect the expression of EMT protein. Clone experiments were performed to detect the proliferation of cells. RNA-seq was further used to analyze the regulatory genes of HOXA5. Using software Cluster 3.0, treeview and DAVID Bioinformatics Resources and Enrichr to analyze KEGG signaling pathway, heatmap. Results: In BT549 cell line, the migration ability of HOXA5 cells was significantly decreased (P <0.001), and the expression of E-cadherin protein was up-regulated. The expression of N-cadherin, Twist1, Slug The protein expression level was significantly down-regulated. The results of plate clone showed that the number of clones formed in the experimental group decreased significantly (P <0.05) and the size of the clone decreased significantly. CONCLUSION: HOXA5 inhibits the migration of breast cancer cells and promotes cell proliferation by promoting the transformation of cells from the stroma to the epithelium. HOXA5 also regulates the proliferation of cancer cells by regulating the pathway of glycolipid metabolism and also inhibits the tumor by regulating cell motility and adhesion-related genes Migration of cells exerts anti-tumor effects via the tumor necrosis factor (TNF) signaling pathway. In conclusion, the transcription factor HOXA5 plays a significant inhibitory role in the development and progression of breast cancer.