摘要
目的探讨真核延伸因子2激酶(eEF2K)基因转染及丹参酮ⅡA磺酸钠(STS)对肺腺癌细胞系A549增殖、侵袭和迁移的影响及其机制。方法预实验筛选STS最佳作用浓度。将A549细胞分为siRNA-NC组(转染siRNA-NC)、siRNA-eEF2K组(转染siRNA-eEF2K)、siRNA-NC+STS组(转染siRNA-NC+10μg/mL STS)和siRNA-eEF2K+STS组(转染siRNA-eEF2K+10μg/mL STS)。CCK-8法、克隆形成实验、Transwell小室法、划痕实验和流式细胞测量术检测细胞存活率、克隆形成率、穿膜细胞数、迁移率和凋亡率;免疫印迹法(Western blot)检测蛋白激酶B(AKT)、磷酸化(p)-AKT、Ki67、基质金属蛋白酶2(MMP-2)和B淋巴细胞瘤-2基因(Bcl-2)蛋白表达。结果与siRNA-NC组比较,siRNA-eEF2K组、siRNA-NC+STS组和siRNA-eEF2K+STS组细胞存活率、克隆形成率、穿膜细胞数、迁移率和p-AKT、Ki67、MMP-2、Bcl-2蛋白均明显降低,细胞凋亡率明显升高(P<0.05),且siRNA-eEF2K+STS组中上述指标变化幅度最大。结论eEF2K沉默联合STS可协同抑制A549细胞增殖,其作用机制可能与抑制p-AKT、Ki67、MMP-2、Bcl-2蛋白表达有关。
Objective To investigate the effects of eukaryotic elongation factor-2 kinase(eEF2K)gene transfection and sodium tanshinoneⅡA 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 10μg/mL STS after siRNA-NC)and siRNA-eEF2K+STS group(treated with 10μg/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.
作者
王布
袁胜芳
张长洪
苑程
黄攀登
张志华
赵建清
WANG Bu;YUAN Sheng-fang;ZHANG Chang-hong;YUAN Cheng;HUANG Pan-deng;ZHANG Zhi-hua;ZHAO Jian-qing(Department of Respiratory Medicine, the First Affiliated Hospital of Hebei Northern College, Zhangjiakou 075000, China)
出处
《基础医学与临床》
2022年第1期106-113,共8页
Basic and Clinical Medicine
基金
河北省卫生厅科研基金(20180818)
张家口市科技攻关计划(1712008D)。
关键词
肺腺癌
真核延伸因子激酶-2
丹参酮ⅡA磺酸钠
增殖
侵袭
lung adenocarcinoma
eukaryotic elongation factor 2 kinase
sodium tanshinoneⅡA sulfonate
proliferation
invasion
