丝氨酸/苏氨酸蛋白激酶1基因沉默对肺癌细胞生物学行为的影响

Effect of small interfering RNA-induced LKB1 gene silencing on the biological behavior of lung carcinoma cells

目的研究丝氨酸/苏氨酸蛋白激酶(LKB1)基因表达沉默对肺癌细胞生物学行为的影响。方法以RNA干扰技术建立LKB1表达抑制的细胞模型,Western blotting方法检测干扰LKB1蛋白表达的效果;通过绘制生长曲线和克隆形成试验检测LKB1基因沉默肺癌细胞的增殖变化;应用Annexin V/PI双染法和流式细胞技术分析LKB1基因沉默后肺癌细胞周期的变化及其凋亡和坏死情况。结果成功建立了LKB1基因沉默肺癌细胞模型,稳定表达LKB1-siRNA细胞的LKB1蛋白表达被明显抑制,LKB1基因沉默后肺癌细胞增殖速度明显加快,但其早期与晚期凋亡率较对照组相比并无显著变化。结论LKB1基因的下调表达对95D细胞部分恶性生物学行为的发生具有一定的促进作用。

Objective To study the effect of the small interfering RNA （siRNA）-mediated LKB1 gene silencing on the biological behavior of lung carcinoma cells. Methods RNA interference technique was used to silence LKB 1 gene in lung carcinoma cells, and the expression of LKB 1 protein were subsequently detected by Western blotting. The cell proliferation was then assessed by observing the growth curve and clone formation of the cells, and cell cycle and apoptosis were assayed by flow cytometry. Results LKB 1 siRNA resulted in remarkable suppression of LKB 1 expression in the lung carcinoma cells. LKB1 gene silencing resulted in accelerated cell proliferation, but cell apoptosis underwent no significant changes. Conclusion Down-regulation of LKB 1 gene expression can stimulate malignant biological behavior of lung carcinoma cells.