沉默LKB1基因对肺癌细胞侵袭迁移和EMT的影响

Effects of LKB1 gene silencing on the invasion, migration and EMT of lung cancer cells

目的探讨沉默肝激酶B1（1iverkinaseB1，LKB1）基因对肺癌95一D和H1299细胞侵袭、迁移能力和上皮间质转化（epithelial to mesenchymal transition，EMT）的影响。方法设计靶向沉默LKB1的shRNA，构建shLKB1重组质粒以及含无关shRNA的shCtd阴性对照质粒，转染肺癌95一D和H1299细胞，Westernblot检测LKB1表达情况，荧光显微镜观察转染效率；Transwell实验和划痕实验检测转染后细胞的侵袭和迁移能力；Western blot检测EMT标志蛋白E一钙黏蛋白（E—cadherin）和波形蛋白（Vimentin）的表达情况。结果成功构建shLKB1重组质粒，并转染人肺癌95-D和H1299细胞。Western blot检测结果显示，与对照组相比，转染组LKB1表达下降，侵袭和迁移能力增强，E—cadhefin表达减少，Vimentin表达增加，差异均具有统计学意义（P〈0．05）。结论沉默抑癌基因LKB1可使肺癌95一D和H1299细胞侵袭和迁移能力增强，并且可能与EMT现象上调有关。

Objective To investigate the effects of liver kinase B1 （LKB1） gene silencing on the invasion, migration and epithelial to mesenchymal transition （EMT） of lung cancer cell lines 95 - D and H1299. Methods A special shRNA was designed to silence LKB1. Then, a recombinant plasmid shLKB1 containing the above shRNA sequence and a negative control plasmid shCtrol containing a unrelated shRNA sequence were constructed, which were then transfected into 95 - D and H1299 cells. The level of LKB1 was measured by Western blotting, while the transfection efficiency was determined by fluorescence microscopy. The invasion and migration capability of the ceils were examined by Transwell and wound - healing assays. The amounts of E - cadherin and Vimentin were detected by Western blotting. Results The recombinant plasmid shLKB1 was successfully constructed and transfected into 95 - D and H1299 cells. Compared with the control, the ceils containing shLKB1 produced reduced levels of LKB1 but increased quantities of E？ cadherin and Vimentin and presented enhanced capacities of invasion and migration （P 〈 0.05 each）. Conclusion LKB1 gene silencing can stimulate 95 - D and H1299 ceils＂ capacities of invasion and migration, which may be associated with enhanced EMT.