Silencing eEF2K combined with sodium tanshinone ⅡA sulfonate synergistically inhibit proliferation of human lung adenocarcinoma cell line A549

Abstract

Objective To investigate the effects of eukaryotic elongation factor-2 kinase (eEF2K) gene transfection and sodium tanshinone Ⅱ A sulfonate (STS) on proliferation, invasion and migration of lung adenocarcinoma cell line A549 and its mechanism. Methods The optimal concentration of STS was screened out by preliminary experiment. A549 cells were divided into siRNA-NC group (transfected with siRNA-NC), siRNA-eEF2K group (transfected with siRNA-eEF2K), siRNA-NC+STS group (treated with 10 μg/mL STS after siRNA-NC) and siRNA-eEF2K+STS group (treated with 10 μg/mL STS after siRNA-eEF2K).The survival rate, clone formation, cell counting of transmembrane cells, migration and apoptosis of A549 cells were detected by CCK-8 method, clone formation test, Transwell chamber test, scratch test and flow cytometry. The protein expression levels of protein kinase B (AKT), phosphorylation (p)-AKT, Ki67, matrix metalloproteinase-2 (MMP-2) and B-cell lymphoma-2 genes (Bcl-2) were detected by Western blot. Results Compared with those in siRNA-NC group, the cell survival rate, clone formation rate, number of transmembrane cells, migration rate and p-AKT, Ki67, MMP-2 and Bcl-2 in proteins siRNA-eEF2K+STS group were significantly lower, while the apoptosis rate was significantly higher (P＜0.05). The siRNA-eEF2K+STS group had the largest range of changes in the above indexes. Conclusions Silencing eEF2K combined with STS can synergistically inhibit the proliferation, invasion and migration of A549 cells and promote cell apoptosis. The mechanism is potentially related to the inhibition of protein expression of p-AKT, Ki67, MMP-2 and Bcl-2.

Keywords

• lung adenocarcinoma|eukaryotic elongation factor 2 kinase|sodium tanshinone ⅱa sulfonate|proliferation|invasion