转染4.1N基因对人肺癌A549细胞株生物学行为的影响

Effect of 4 . 1N gene transfection on biological characteristics of human lung cancer cell line ;A549

目的：探讨4.1N在肺癌A549细胞株中的表达以及细胞转染前后增殖、侵袭、迁移等的变化。方法脂质体介导真核表达载体pEGFP-4.1N转染人肺腺癌细胞株A549，同时设空载体组及未转染组为对照，48 h后用半定量反转录-聚合酶链反应（RT-PCR）和Western blot方法检测各组细胞4.1N在mRNA和蛋白水平的表达；通过四甲基偶氮唑盐（MTT）法测定各组细胞的增殖能力；Transwell小室模型检测各组细胞的体外侵袭、迁移能力。结果 A549转染pEGFP-4.1N 48 h后，4.1N的mRNA及蛋白表达水平均提高（P〈0.05），细胞增殖活性显著降低（P〈0.05），细胞的体外迁移及侵袭能力明显减弱（P〈0.05）。结论转染4.1N基因能显著上调人高转移肺腺癌A549细胞株中4.1N mRNA和蛋白的表达水平。4.1N基因可以抑制A549细胞株的增殖、黏附、侵袭和迁移，在肺癌转移中起重要作用，有可能成为肺癌转移的分子标志物。

Objective To investigate the effects of 4.1N expression in lung cancer A549 cell line on cell proliferation, invasion and migration. Methods A549 cells were cultured in vitro and transfected with lipofectamine 2000 mediation. Three groups were employed： transfection with pEGFP-4.1N plasmid, pEGFP vector plasmid, and blank control, respectively. The mRNA and protein expression differences of 4.1N was examined by semi-quantitative RT-PCR and Western blot in every group after 48 h. The proliferation capability was determined by MTT assay. Invasion capability was evaluated by scratches, adhesion experiments and Transwell chamber model. Results After the transfection, the expression of 4.1N mRNA and protein in pEGFP-4.1N plasmid transfection group was significantly enhanced （P〈0.05）. The proliferation capability of A549 cells descended extremely （P〈0.05）. The migration and invasion capability of A549 cells in vitro decreased substantially （P〈0.05）. Conclusions Transfected with 4.1N gene can significantly increases the expression levels of 4.1N mRNA and protein in A549 cells which are highly metastatic in human. Cell behavior in vitro studies showed that 4.1N gene can inhibit the proliferation, adhesion, invasion and migration of A549 cells, which plays an important role in the metastasis of lung cancer and it may become a molecular marker for metastasis of lung cancer.