摘要
目的 探究阿美替尼联合安罗替尼抑制非小细胞肺癌的作用.方法 CCK-8法、集落克隆法和流式细胞术检测不同浓度阿美替尼和安罗替尼对PC-9细胞和HCC827细胞增殖、细胞存活和细胞凋亡的影响;SynergyFinder模型评价阿美替尼与安罗替尼的协同作用;Transwell小室实验检测阿美替尼联合安罗替尼对PC-9细胞和HCC827细胞侵袭及迁移的作用;Western blotting实验检测阿美替尼联合安罗替尼对PC-9细胞和HCC827细胞凋亡和侵袭迁移相关蛋白Bax、Bcl-2、E-Cadherin、Vimentin、MMP2和MMP9及PI3K-Akt通路关键蛋白表达的影响.结果 阿美替尼作用于PC-9细胞的IC50为1.701 μmol/L,安罗替尼作用于PC-9细胞的IC50=4.979 μmol/L,协同得分(ZIP)为19.112;阿美替尼对HCC827细胞IC50=2.961 μmol/L,安罗替尼对HCC827细胞IC50=7.934 μmol/L,协同得分(ZIP)为12.325,阿美替尼联合安罗替尼能够显著抑制PC-9细胞和HCC827细胞增殖(P<0.05).相比于单药组,集落克隆结果表明阿美替尼联合安罗替尼能够显著抑制PC-9细胞和HCC827细胞存活(P<0.01);流式细胞术结果表明,阿美替尼联合安罗替尼能够促进PC-9细胞和HCC827细胞发生凋亡(P<0.05);Transwell实验结果表明,阿美替尼联合安罗替尼能够显著抑制PC-9细胞和HCC827细胞的侵袭迁移能力(P<0.01);Western blotting实验结果表明,阿美替尼联合安罗替尼能够显著促进PC-9细胞和HCC827细胞中E-Cadherin和Bax蛋白的表达,抑制Bcl-2、Vimentin、MMP2和MMP9蛋白的表达(P<0.01),同时显著降低了PI3K-Akt通路中关键蛋白PI3K、Akt的磷酸化蛋白水平(P<0.01).结论 阿美替尼联合安罗替尼通过下调PI3K-Akt通路抑制NSCLC肿瘤细胞,这种联合治疗方式可能成为临床上治疗 NSCLC患者的一种潜在治疗策略.
Abstract
Objective To investigate the inhibitory effect of aumolertinib combined with anlotinib on proliferation of non-small cell lung cancer(NSCLC)cells.Methods CCK-8 assay,colony formation assay,and flow cytometry were used to assess the effect of different concentrations of aumolertinib or anlotinib on proliferation,survival,and apoptosis of PC-9 and HCC827 cells,and their synergistic effect was evaluated using the SynergyFinder model.In PC-9 and HCC827 cells treated with aumolertinib combined with anlotinib,the changes in cell invasion and migration abilities were assessed with Transwell assay,and the expressions of apoptosis-and invasion/migration-related proteins(Bax,Bcl-2,E-cadherin,vimentin,MMP2,and MMP9)and the key PI3K-Akt pathway proteins were detected using Western blotting.Results In PC-9 cells,the IC50 of aumolertinib and anlotinib was 1.701 μmol/L and 4.979 μmol/L,respectively,with a synergy score(ZIP)of 19.112;in HCC827 cells,their IC50 was 2.961 μmol/L and 7.934 μmol/L,respectively,with a ZIP of 12.325.Compared with aumolertinib and anlotinib used alone,their combined treatment more strongly inhibited the proliferation and survival,enhanced apoptosis and suppressed invasion and migration abilities of PC-9 and HCC827 cells.Western blotting showed that in both PC-9 and HCC827 cells,the combined treatment significantly upregulated the expressions of E-cadherin and Bax proteins,downregulated the expressions of Bcl-2,vimentin,MMP2,and MMP9 proteins,and reduced phosphorylation levels of PI3K and Akt.Conclusion Aumolertinib combined with anlotinib can effectively inhibit NSCLC cell proliferation by downregulating the PI3K-Akt pathway,suggesting a potentially new option for NSCLC treatment.
基金项目
安徽省重点研究与开发计划项目(202104g01020017)
安徽省重点生化工程中心科研平台开放课题(2023SYKFD05)
国家级大学生创新创业训练计划项目(202210367064)
安徽省大学生创新创业训练计划项目(S202310367111)
安徽省优秀科研创新团队项目(2022AH010084)
安徽省自然科学基金(1908085QH373)
安徽高校自然科学研究项目(KJ2020A0565)
安徽高校自然科学研究项目(KJ2021A0702)
维普
万方数据