LEF1慢病毒表达载体的构建及稳定肺癌细胞株的鉴定

Construction of LEF1-overexpressed Lentiviral Vector and Identification of Stable LEF1-overexpressed Lung Cancer Cells

肺癌作为恶性肿瘤死亡的主要原因,其发生机制涉及诸多肿瘤相关基因的异常表达。淋巴增强子结合因子1(LEF1)是经典Wnt/β-catenin信号通路的关键转录因子,在多种肿瘤的侵袭及转移过程中发挥重要作用。本文通过构建LEF1过表达的慢病毒载体,在293T细胞中包装慢病毒颗粒,感染目的肺癌细胞并筛选获得稳定表达LEF1的A549及H1299肺癌细胞株,为后续鉴定LEF1直接调控的靶基因奠定细胞学基础。功能研究结果显示,过表达LEF1使上皮细胞标志物E-cadherin表达下调,而间充质细胞标志物N-cadherin表达上调,因而对肺癌细胞的上皮-间充质转化过程(Epithelial-mesenchymal transition,EMT)具有明显的促进作用。

As the leading cause of cancer death, lung cancer develops with a complexed mechanism which involves in the aberrant expressions of various cancer related genes. Lymphoid enhancer-binding factor 1（LEF1）, which is a critical transcription factor of canonical Wnt/β-catenin signaling pathway, plays important roles in the process of tumor invasion and metastasis. In this study, LEF1-overexpressed lentiviral vector was constructed and transfected into HEK-293 T cells for virus packaging. A549 and H1299 cells were infected with recombinant lentivirus and screened for stable LEF1-overexpressed lung cancer cells, on the basis of which direct-regulated target genes of LEF1 can be subsequently identified. As a functional result, forced expression of LEF1 reduced the epithelial marker, E-cadherin, and increased the mesenchymal marker, N-cadherin, which exerted a promotion effect on the epithelial-mesenchymal transition（EMT） of lung cancer cells.