摘要
目的探究安罗替尼对人肺鳞癌NCI-H226细胞增殖、迁移、侵袭和上皮间质转化(EMT)的影响及其机制。方法将人肺鳞癌NCI-H226细胞分为对照组(A组)、不同浓度(10、20、40μmol/L)安罗替尼处理组(B~D组)、安罗替尼(20μmol/L)+PI3K/AKT信号通路抑制剂LY294002(30μmol/L)处理组(E组)和安罗替尼(20μmol/L)+PI3K/AKT信号通路激活剂SC79(10μmol/L)处理组(F组)。通过CCK-8法检测各组细胞的细胞活力,通过划痕实验检测各组细胞的迁移率,通过Transwell侵袭实验检测各组细胞的侵袭能力,通过RT-qPCR法检测各组细胞中EMT相关基因E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)和纤维粘连蛋白(FN)mRNA的相对表达水平,通过蛋白免疫印迹(WB)法检测各组细胞中PI3K、AKT、p-PI3K、p-AKT、E-cadherin、N-cadherin、Vimentin和FN蛋白的相对表达水平。结果实验结果显示,与A组相比,B~D组细胞活力、迁移率、侵袭细胞数及p-PI3K和p-AKT蛋白的相对表达水平均显著下降,且呈剂量依赖性(F=49.315~145.576,t_(LSD)=2.459~251.731,P<0.05)。与A组相比,C组细胞中N-cadherin、Vimentin、FN mRNA和蛋白的相对表达水平下降,E-cadherin mRNA和蛋白相对表达水平显著升高(F=30.554~136.286,t_(LSD)=4.158~8.315,P<0.05);与C组相比,E组细胞活力、迁移率、侵袭能力、p-PI3K和p-AKT蛋白相对表达水平以及N-cadherin、Vimentin、FN mRNA和蛋白的相对表达水平显著降低,F组上述指标显著升高(t_(LSD)=3.221~11.079,P<0.05);E组细胞中E-cadherin mRNA和蛋白的相对表达水平显著高于C组,F组上述指标显著低于C组(t_(LSD)=2.195~7.213,P<0.05)。结论安罗替尼可抑制NCI-H226细胞的增殖、迁移、侵袭和EMT,这一过程可能与抑制PI3K/AKT信号通路活化有关。
Objective To investigate the effect of anlotinib on the proliferation,migration,invasion,and epithelial-me-senchymal transition(EMT)of human lung squamous cell carcinoma NCI-H226 cells.Methods Human lung squamous cell carcinoma NCI-H226 cells were divided into control group(group A),anlotinib treatment groups at different concentrations(10,20,and 40μmol/L)(groups B-D),anlotinib(20μmol/L)+PI3K/AKT signaling pathway inhibitor LY294002(30μmol/L)treatment group(group E),and anlotinib(20μmol/L)+PI3K/AKT signaling pathway activator SC79(10μmol/L)treatment group(group F).CCK-8 assay was used to measure cell viability;scratch assay was used to measure the migration rate of cells;Transwell assay was used to measure the invasion ability of cells;RT-qPCR was used to measure the relative mRNA expression levels of the EMT-related genes E-cadherin,N-cadherin,Vimentin,and fibelectin(FN)in cells,and Western blotting was used to measure the relative protein expression levels of PI3K,AKT,p-PI3K,p-AKT,E-cadherin,N-cadherin,Vimentin,and FN in cells.Results The results of the experiment showed that compared with group A,groups B-D had significant reductions in cell viability,cell migration rate,the number of invasive cells,and the relative protein expression levels of p-PI3K and p-AKT(F=49.315-145.576,t_(LSD)=2.459-251.731,P<0.05).Compared with group A,group C had significant reductions in the relative mRNA and protein expression levels of N-cadherin,Vimentin,and FN and significant increases in the mRNA and protein expression levels of E-cadherin(F=30.554-136.286,t_(LSD)=4.158-8.315,P<0.05).Compared with group C,group E had significant reductions in cell viability,cell migration rate,invasion ability,the relative protein expression levels of p-PI3K and p-AKT,and the relative mRNA and protein expression levels of N-cadherin,Vimentin,and FN,and group F had significant increases in the above indicators(t_(LSD)=3.221-11.079,P<0.05).Compared with group C,group E had significantly higher relative mRNA and protein expression levels of E-cadherin,while group F had significantly lower expression levels than group C(t_(LSD)=2.195-7.213,P<0.05).Conclusion Anlotinib can inhibit the proliferation,migration,invasion,and EMT of human lung squamous cell carcinoma NCI-H226 cells,possibly by inhibiting the activation of the PI3K/AKT signaling pathway.
作者
赵振波
马胜喜
刘德义
ZHAO Zhenbo;MA Shengxi;LIU Deyi(Department of Respiratory and Critical Me-dicine,Xinxiang Central Hospital,Xinxiang 453000,China)
出处
《精准医学杂志》
2026年第1期18-24,共7页
Journal of Precision Medicine
基金
新乡市中心医院新技术项目[2021-023-01(K)]。
