摘要
探究安罗替尼干预对人肺鳞癌NCI-H226细胞糖酵解、增殖、克隆形成、迁移的影响及作用机制,为肺癌的体外试验提供参考依据.将人肺鳞癌NCI-H226细胞分为对照组(不做干预)、10、20、40 μmol/L安罗替尼组和阳性药物组(20 μg/mL顺铂).用活细胞计数(CCK-8)、5-乙炔基-2′脱氧尿嘧啶核苷(EdU)、平板克隆形成、乳酸检测试剂盒、葡萄糖检测试剂盒、Transwell小室、实时荧光定量PCR(RT-qPCR)及蛋白免疫印迹(Western blotting)法对增殖活性、增殖率、克隆形成率、乳酸含量、葡萄糖消耗水平、细胞迁移数及上皮细胞间质化(EMT)相关因子表达水平进行测定.对照组、10、20、40 μmol/L安罗替尼组、阳性药物组NCI-H226细胞增殖活性、增殖率、克隆形成率、乳酸含量、葡萄糖消耗水平及N-钙黏蛋白(N-cadherin)、波形蛋白(vimentin)和纤维黏连蛋白(FN)mRNA和蛋白表达水平组间比较差异具有统计学意义(P<0.05).20、40 μmol/L安罗替尼和阳性药物组细胞增殖活性高于对照组(P<0.05).10、20、40 μmol/L安罗替尼组和阳性药物组细胞增殖率、克隆形成率、乳酸含量、葡萄糖消耗水平及N-cadherin、vimentin和FN mRNA和蛋白表达水平高于对照组(P<0.05).安罗替尼可显著抑制人肺鳞癌NCI-H226细胞的糖酵解、增殖、克隆形成及迁移.
Abstract
To investigate the effects and mechanism of anlotinib intervention on glycolysis,proliferation,colony formation and migration of human lung cancer NCI-H226 cells,to provide reference for in vitro test of lung cancer.Human lung cancer NCI-H226 cells were divided into control group which is without intervention,experimental group involving 10,20,40 μmol/L anlotinib,and positive control group involving 20 μg/mL cisplatin.The cell counting kit-8(CCK-8),5-acetylene-2′ deoxyura-cil riboside(EdU)staining,plate clone formation,lactic acid detection kit,glucose detection kit,Transwell assay,RT-qPCR and Western blotting(WB)assays were used to detect proliferation viability,proliferation,colony-formation ability,lactic acid con-tent,glucose consumption level,cell migration number and expression level of related factor levels.The proliferation activity,proliferation rate,clone formation rate,lactate content,glucose consumption level,N-cadherin,vimentin and fibronectin(FN)of NCI-H226 cells in control group,experimental group and positive drug group.There were significant differences in mRNA and protein expression levels between groups(P<0.05).The cell proliferation activity of 20 and 40 μmol/L anlotinib and positive drug groups was higher than that of control group(P<0.05).The cell proliferation rate,clone formation rate,lactate content,glucose consumption level and mRNA and protein expression levels of N-cadherin,vimentin and FN in the experimental group and the positive drug group were higher than those in the control group(P<0.05).Anlotinib can significantly inhibit glycolysis,proliferation,colony formation and migration of human lung cancer NCI-H226 cells.
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