eEF2K沉默联合丹参酮ⅡA磺酸钠协同抑制人肺腺癌细胞系A549增殖

doi:10.16352/j.issn.1001-6325.2022.01.017

基础医学与临床 ›› 2022, Vol. 42 ›› Issue (1): 106-113.doi: 10.16352/j.issn.1001-6325.2022.01.017

eEF2K沉默联合丹参酮ⅡA磺酸钠协同抑制人肺腺癌细胞系A549增殖

王布, 袁胜芳, 张长洪, 苑程, 黄攀登, 张志华, 赵建清^*

河北北方学院附属第一医院呼吸内科, 河北张家口 075000

收稿日期:2020-11-09 修回日期:2021-05-15 出版日期:2022-01-05 发布日期:2022-01-05
通讯作者: ^* whkj86@163.com
基金资助:
河北省卫生厅科研基金(20180818);张家口市科技攻关计划(1712008D)

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

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

Received:2020-11-09 Revised:2021-05-15 Online:2022-01-05 Published:2022-01-05
Contact: ^* whkj86@163.com

摘要/Abstract

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+10 μg/mL STS)和siRNA-eEF2K+STS组(转染siRNA-eEF2K+10 μg/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。

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

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.

Key words: lung adenocarcinoma, eukaryotic elongation factor 2 kinase, sodium tanshinone ⅡA sulfonate, proliferation, invasion

中图分类号:

R734.2

王布, 袁胜芳, 张长洪, 苑程, 黄攀登, 张志华, 赵建清. eEF2K沉默联合丹参酮ⅡA磺酸钠协同抑制人肺腺癌细胞系A549增殖[J]. 基础医学与临床, 2022, 42(1): 106-113.

WANG Bu, YUAN Sheng-fang, ZHANG Chang-hong, YUAN Cheng, HUANG Pan-deng, ZHANG Zhi-hua, ZHAO Jian-qing. Silencing eEF2K combined with sodium tanshinone ⅡA sulfonate synergistically inhibit proliferation of human lung adenocarcinoma cell line A549[J]. Basic & Clinical Medicine, 2022, 42(1): 106-113.

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eEF2K沉默联合丹参酮ⅡA磺酸钠协同抑制人肺腺癌细胞系A549增殖

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

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