抑癌基因Lkb1超表达载体构建及其在人舌鳞癌细胞SCC-15中的生物活性分析

目的:了解抑癌基因LKB1对人舌鳞癌细胞的影响,初步探讨其相关通路机制。方法:构建LKB1真核表达载体,通过脂质体转染人舌鳞癌细胞系(SCC-15),利用RT-PCR、Western blot 和流式细胞术来检测SCC-15中基因表达和细胞周期凋亡的变化。结果:LKB1在SCC-15细胞中mRNA水平上成功过表达约300倍,蛋白表达水平也显著上调;同时发现LKB1超表达对细胞周期起阻滞作用,并可上调p53、PTEN和TGF-β,同时下调VEGF的表达,差异均有统计学意义。结论:成功构建了LKB1真核表达载体,该基因在SCC-15细胞中超表达对细胞周期起阻滞作用,调控p53、PTEN、TGF-β和VEGF基因表达,可能参与调控p53和TGF-β等多种信号通路,为进一步研究 LKB1 基因在恶性舌鳞癌发病的机制和临床治疗的作用提供了重要实验依据。

Construction of LKB1 Overexpression Vector and Its Biological Activity in Human Tongue Squamous Cell Carcinoma SCC-15 Cells

Objective: To investigate the regulatory role of LKB1 gene and related signaling pathway in human tongue squamous cell carcinoma cells. Methods: Eukaryotic expression vector of LKB1 gene was constructed, and transfected into SCC-15 cells. RT-PCR, western blot and flow cytometry were used to detect the expression of related molecules and the effects on cell cycle and apoptosis. Results: The expression of LKB1 was significantly increased in SCC-15 cells after transfected. Meanwhile, cell cycle arrest was induced by over-expression of LKB1. In addition, the expression of p53, TGF-β and PTEN was up-regulated, while the expression of VEGF was down-regulated. Conclusion: Eukaryotic expression vector of LKB1 gene was successfully constructed. Additionally, LKB1 could regulate cell cycle and the expression of p53, PTEN, TGF-β and VEGF in human tongue squamous cell carcinoma cells.