槲皮素对人肺癌A549细胞移植瘤模型小鼠移植瘤生长和肺转移及细胞侵袭和迁移的影响及其机制  

Effects of quercetin on growth and lung metastasis of transplanted tumor and cell invasion,and cell migration in human lung cancer A549 cells transplanted tumor model mice and their mechanisms

作　　者：靳莉 张小红 胡朝阳 李凤芝 崔永亮 李杨 刘倩倩 乔艳俊 JIN Li;ZHANG Xiaohong;HU Chaoyang;LI Fengzhi;CUI Yongliang;LI Yang;LIU Qianqian;QIAO Yanjun(Department of Respiratory and Critical Care Medicine,Affiliated Zhengzhou Central Hospital,Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学附属郑州中心医院呼吸与危重症医学科,河南郑州450000

出　　处：《吉林大学学报（医学版）》2023年第4期1018-1026,共9页Journal of Jilin University：Medicine Edition

基　　金：河南省科技厅科技攻关项目(182102310314)。

摘　　要：目的:探讨槲皮素(QUE)对人肺癌A549细胞移植瘤模型小鼠移植瘤生长、肺转移、细胞侵袭和迁移的影响,并阐明其相关机制。方法:小鼠分为对照组(无干预)、阳性药物组(5.0 mg·kg^(-1)顺铂)、低剂量(12.5 mg·kg^(-1))QUE组、中剂量(25.0 mg·kg^(-1))QUE组和高剂量(50.0 mg·kg^(-1))QUE组,除对照组外其余各组小鼠采用A549细胞制备移植瘤模型,治疗10 d后计算各组小鼠肿瘤质量抑制率和肺转移抑制率。体外培养人肺癌A549细胞,分为正常对照组、QUE组、QUE+Runt相关转录因子3(Runx3)阴性对照序列(si-NC)(QUE+si-NC)组和QUE+Runx3小干扰序列(si-Runx3)(QUE+si-Runx3)组。实时荧光定量PCR(RT-qPCR)法检测各组A549细胞中Runt mRNA表达水平,CCK-8法检测各组细胞增殖抑制率,Transwell小室实验检测各组A549细胞中侵袭细胞数和迁移细胞数,Western blotting法检测各组小鼠移植瘤组织及各组A549细胞中Runx3、Wnt3a、β-连环蛋白(β-catenin)、基质金属蛋白酶2(MMP-2)和基质金属蛋白酶9(MMP-9)蛋白表达水平。结果:与对照组比较,阳性药物组及低、中和高剂量QUE组小鼠肿瘤质量抑制率、肺转移抑制率和移植瘤组织中Runx3蛋白表达水平均明显升高(P<0.05),移植瘤组织中Wnt3a、β-catenin、MMP-2和MMP-9蛋白表达水平均明显降低(P<0.05)。与正常对照组比较,QUE组A549细胞增殖抑制率、A549细胞中Runx3 mRNA及蛋白表达水平均明显升高(P<0.05),侵袭细胞数和迁移细胞数明显减少(P<0.05),A549细胞中Wnt3a、β-catenin、MMP-2和MMP-9蛋白表达水平均明显降低(P<0.05);与QUE+si-NC组比较,QUE+si-Runx3组A549细胞增殖抑制率、A549细胞中Runx3mRNA及蛋白表达水平均明显降低(P<0.05),侵袭细胞数和迁移细胞数明显增加(P<0.05),A549细胞中Wnt3a、β-catenin、MMP-2和MMP-9蛋白表达水平均明显升高(P<0.05)。结论:QUE可抑制人肺癌A549细胞移植瘤小鼠移植瘤转移和侵袭,其机制可能是通过促进Runx3表达抑制Objective:To detect the effect of quercetin(QUE)on the growth,lung metastasis,cell invasion and migration of transplanted tumor of the human lung cancer A549 cells transplanted tumor in the mice,and to clarify its related mechanism.Methods:The mice were divided into control group(no intervention),positive drug group(5 mg·kg^(-1)cisplatin)and low,medium and high doses(12.5,25.0,and 50.0 mg·kg^(-1) QUE)of QUE groups.Except for control group,the mice in other groups were constructed transplanted tumor models using A549 cells.The inhibitory rates of tumor weights and inhibitory rates of lung metastasis of mice in various groups were detected after treated for 10 d.The human lung cancer A549 cells were cultured in vitro and divided into normal control group,QUE group,QUE+Runt related transcription factor 3(Runx3)negative control sequence(si-NC)(UE+si-NC)group,and QUE+Runx3 interference sequence(si-Runx3)(QUE+si-Runx3)group.Real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of Runt mRNA in the A549 cells in various groups;CCK-8 assay was used to detect inhibitory rates of proliferation of cells in various groups;Transwell chamber experiment was used to detect the numbers of invasion and migration cells in various groups;Western blotting method was used to detect the expression levels of Runx3,Wnt3a,β-catenin,matrix metallopeptidase-2(MMP-2),and matrix metallopeptidase-9(MMP-9)proteins in cells in various groups.Results:Compared with control group,the inhibitory rate of tumor weight,inhibitory rate of lung metastasis and the expression levels of Runx3 protein in transplant tumor tissue of the mice in positive drug group,low,medium and high doses of QUE groups were increased(P<0.05),and the expression levels of Wnt3a,β-catenin,MMP-2,and MMP-9 proteins were decreased(P<0.05).Compared with normal control group,the inhibitory rate of proliferation of the A549 cells in QUE group,the expression levels of Runx3 mRNA and protein in the A549 cells in QUE group were significantly i

关 键 词：槲皮素 肺肿瘤 A549细胞 细胞侵袭 细胞迁移 RUNT相关转录因子3 Wnt/β-catenin信号通路 

分 类 号：R734.2[医药卫生—肿瘤]