安罗替尼对MYCN扩增神经母细胞瘤增殖凋亡与迁移侵袭的影响及机制研究  

Effects and mechanisms of anlotinib on proliferation,apoptosis,migration,and invasion in MYCN-amplified neuroblastomas

作　　者：苏雨栋 赵强 Yudong Su;Qiang Zhao(Department of Thoracic Oncology,Tianjin Medical University Cancer Institute&Hospital,National Clinical Research Center for Cancer,State Key Laboratory of Druggability Evaluation and Systematic Translational Medicine,Tianjin Key Laboratory of Cancer Prevention and Therapy,Tianjin's Clinical Research Center for Cancer,Tianjin Lung Cancer Center,Tianjin 300060,China;Department of Pediatric Oncology,Tianjin Medical University Cancer Institute&Hospital,National Clinical Research Center for Cancer,State Key Laboratory of Druggability Evaluation and Systematic Translational Medicine,Tianjin Key Laboratory of Cancer Prevention and Therapy,Tianjin's Clinical Research Center for Cancer,Tianjin Lung Cancer Center,Tianjin 300060,China)

机构地区：[1]天津医科大学肿瘤医院肺部肿瘤内科,国家肿瘤临床医学研究中心,药物成药性评价与系统转化全国重点实验室,天津市“肿瘤防治”重点实验室,天津市恶性肿瘤临床医学研究中心,天津市肺癌诊治中心 [2]天津医科大学肿瘤医院肺部肿瘤内科,国家肿瘤临床医学研究中心,药物成药性评价与系统转化全国重点实验室,天津市“肿瘤防治”重点实验室,天津市恶性肿瘤临床医学研究中心,天津市肺癌诊治中心,儿童肿瘤科

出　　处：《中国肿瘤临床》2024年第24期1250-1254,共5页Chinese Journal of Clinical Oncology

基　　金：天津市教委科研计划项目(编号:2023KJ081)资助。

摘　　要：目的:探讨安罗替尼对MYCN扩增神经母细胞瘤增殖、凋亡、迁移与侵袭的影响及机制。方法:采用不同浓度的安罗替尼作用于2种MYCN扩增NB细胞系,利用CCK8法和平板克隆实验检测细胞增殖能力;流式细胞术检测细胞凋亡情况;划痕实验测定细胞迁移能力;Transwell实验检测细胞侵袭能力;Western blot实验检测信号通路关键蛋白的表达情况。结果:CCK8结果显示安罗替尼对MYCN扩增NB细胞系的抑制呈浓度和时间依赖性,平板克隆实验结果显示安罗替尼显著抑制细胞克隆形成(P<0.0001);划痕实验结果显示安罗替尼显著降低MYCN扩增NB细胞系的迁移能力(P<0.0001);Transwell实验结果显示安罗替尼显著抑制MYCN扩增NB细胞系的侵袭能力(P<0.0001);凋亡实验结果显示安罗替尼显著促进MYCN扩增NB细胞系发生细胞凋亡(P<0.0001);Western blot结果显示安罗替尼显著降低MYCN扩增NB的VEGFR2和AKT蛋白磷酸化水平,其下游关键蛋白N-cadherin和Bcl-2的表达显著下调,E-cadherin和Bax的表达显著上调。结论:安罗替尼通过VEGFR2/PI3K/AKT信号通路显著抑制NB增殖、迁移和侵袭,促使其凋亡。Objective:To study the effects and mechanisms of anlotinib on the proliferation,migration,invasion,and apoptosis of MYCNamplified neuroblastomas(NB).Methods:Two MYCN-amplified NB cell lines were treated with varying concentrations of anlotinib.CCK8 and clone formation assays were employed to detect cell proliferation,flow cytometry was used to detect cell apoptosis,scratch tests were performed to determine cell migration,and Transwell assays were used to detect cell invasion.Results:CCK8 assays indicated that anlotinib inhibited proliferation in MYCN-amplified NB cell lines in a concentration-and time-dependent manner.Clone formation assays showed that anlotinib significantly inhibited cell clone formation(P<0.0001).The results of the apoptosis experiment showed that anlotinib significantly promoted apoptosis in MYCN-amplified NB cell lines(P<0.0001).Scratch assays showed that anlotinib significantly reduced the migration ability of MYCN-amplified NB cells(P<0.0001).Transwell assays revealed that anlotinib significantly inhibited the invasive abilities of MYCNamplified NB cell lines(P<0.0001).Western blot analysis demonstrated that anlotinib significantly reduced the phosphorylation of VEGFR2 and AKT proteins in MYCN-amplified NB cells.The expression of the key downstream proteins N-cadherin and Bcl-2 was significantly downregulated,whereas the expression of E-cadherin and Bax was significantly upregulated.Conclusions:Anlotinib markedly inhibits the proliferation,migration,and invasion of MYCN-amplified NB cells and induces apoptosis via the VEGFR2/PI3K/AKT signaling pathway.

关 键 词：安罗替尼 神经母细胞瘤 MYCN扩增 细胞增殖 细胞凋亡 

分 类 号：R739.4[医药卫生—肿瘤]