仑伐替尼对甲状腺癌TPC-1细胞生物学行为的影响  

Effects of lenvatinib on biological behavior of thyroid cancer TPC-1 cells

作　　者：齐蕾[1] 叶健文[2] 薛文华[1] 张会娟[3] QI Lei;YE Jianwen;XUE Wenhua;ZHANG Huijuan(Department of Pharmacy,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Hepatobiliary and Pancreatic Surgery,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052;Department of Endocrinology,the First Affiliated Hospital,Zhengzhou University,Zhengzhou 450052)

机构地区：[1]郑州大学第一附属医院药学部,郑州450052 [2]郑州大学第一附属医院肝胆胰外科,郑州450052 [3]郑州大学第一附属医院内分泌科,郑州450052

出　　处：《郑州大学学报（医学版）》2022年第2期261-266,共6页Journal of Zhengzhou University(Medical Sciences)

基　　金：河南省医学科技攻关计划联合共建项目(LHGJ20190028);河南省高等学校重点科研项目(19B320039)。

摘　　要：目的:探讨仑伐替尼对甲状腺癌TPC-1细胞生物学行为的影响。方法:TPC-1细胞分为阴性对照组(不处理)与仑伐替尼组(20μmol/L仑伐替尼处理),采用克隆形成实验和Annexin V-FITC/PI双染法检测细胞克隆和凋亡情况;TPC-1细胞分为阴性对照组(不处理)与仑伐替尼组(40μmol/L仑伐替尼处理),应用Transwell小室检测细胞转移和侵袭情况;TPC-1细胞分为阴性对照组(不处理)与20、40μmol/L仑伐替尼组,应用Western blot法检测细胞Cleaved Caspase-9、基质金属蛋白酶2(MMP-2)、磷酸化的蛋白激酶B(p-AKT)、LC3Ⅱ、LC3Ⅰ、P62蛋白表达;TPC-1细胞分为阴性对照组(不处理)、仑伐替尼组(20μmol/L仑伐替尼处理)、3-甲基腺嘌呤(3-MA)组(5 mmol/L 3-MA处理)、仑伐替尼+3-MA组,采用Western blot法检测LC3Ⅰ、LC3Ⅱ蛋白的表达,采用CCK-8法检测细胞增殖情况,采用Annexin V-FITC/PI双染法检测细胞凋亡情况。10只裸鼠于右侧腋下接种约1×10^(7)个TPC-1细胞建立移植瘤模型,待肿瘤生长至平均体积100 mm^(3)时分为对照组(胃内灌注PBS)和仑伐替尼组[胃内灌注仑伐替尼25 mg/(kg·d)],比较3周后瘤体体积和质量。结果:与阴性对照组比较,仑伐替尼组细胞克隆数减少、凋亡率增加、侵袭和转移能力减弱;仑伐替尼组TPC-1细胞Cleaved Caspase-9蛋白表达水平和LC3Ⅱ/LC 3Ⅰ比值升高,MMP-2、p-AKT、P62蛋白表达水平降低(P<0.05)。仑伐替尼联合3-MA增强TPC-1细胞的生长抑制和凋亡(P<0.05)。仑伐替尼组裸鼠移植瘤体积及质量较对照组降低(P<0.05)。结论:仑伐替尼抑制TPC-1细胞生长及转移、诱导凋亡及自噬,可能与p-AKT/Cleaved Caspase-9/MMP-2通路有关。Aim:To explore the effects of lenvatinib on biological behavior of thyroid cancer TPC-1 cells.Methods:TPC-1 cells were divided into negative control group and lenvatinib group(treated with 20μmol/L lenvatinib).The colony ability and apoptosis of TPC-1 cells were detected by colony formation assay and Annexin V-FITC/PI double staining.TPC-1 cells were divided into negative control group and lenvatinib group(treated with 40μmol/L lenvatinib).The invasion and migration of TPC-1 cells were detected by Transwell assay.TPC-1 cells were divided into negative control group and lenvatinib group(treated with 20 and 40μmol/L lenvatinib).Western blot was used to detect the expression of Cleaved Caspase-9,matrix metalloproteinase 2(MMP-2),phospho-AKT kinase(p-AKT),LC3Ⅱ,LC3Ⅰ,P62.TPC-1 cells were divided into negative control group,lenvatinib group(treated with 20μmol/L lenvatinib),3-methyladenine(3-MA)group(treated with 5 mmol/L 3-MA)and lenvatinib combined with 3-MA group.The expression of LC3Ⅱ,LC3Ⅰwere detected by Western blot.The proliferation and apoptosis of TPC-1 cells were detected by CCK-8 assay and Annexin V-FITC/PI double staining,respectively.Ten nude mice were inoculated with 1×10^(7) TPC-1 cells into right flank to establish the transplanted tumor model.When the average volume of tumor reached 100 mm^(3),the mice were divided into control group(received PBS,intragastric administration),and lenvatinib group[received lenvatinib,25 mg/(kg·d),intragastric administration].The tumor volume and weight were measured and compared after 3 weeks.Results:Compared with negative control group,the colony ability of TPC-1 cells treated with lenvatinib was reduced,invasion and migration abilities decreased,while the apoptosis rate of TPC-1 cells treated with lenvatinib was higher;treatment with lenvatinib increased the protein expression levels of Cleaved Caspase-9 and LC3Ⅱ/LC3Ⅰratio,whereas decreased expressions of MMP-2,p-AKT and P62(P<0.05).The relative cell viability in lenvatinib combined with 3-MA group was

关 键 词：甲状腺癌 仑伐替尼 自噬 凋亡 p-AKT通路 TPC-1细胞 

分 类 号：R736.1[医药卫生—肿瘤]