miR-363-3p对非小细胞肺癌细胞迁移与侵袭能力的影响  

Effects of miR-363-3p on migration and invasion of non-small-cell lung cancer

作　　者：殷玉 王亚红 许志亮 刘刚 YIN Yu;WANG Yahong;XU Zhiliang;LIU Gang(Respiratory Medicine Department,Affiliated Hospital of Guangdong Medical University,Zhanjiang 524000,China;不详)

机构地区：[1]广东医科大学附属医院呼吸内科,524000 [2]广东医科大学附属医院临床研究中心

出　　处：《山东医药》2021年第23期14-17,共4页Shandong Medical Journal

基　　金：国家自然科学基金面上项目(81570062、82070061);广东省医学计划项目(A2019456);广东医科大学附属医院科研项目(GDMUM201801)。

摘　　要：目的观察miR-363-3p对非小细胞肺癌(NSCLC)细胞迁移与侵袭能力的影响。方法收集发生和未发生淋巴结转移的NSCLC组织样本各26、21例份,常规培养人NSCLC细胞株A549及高转移性NSCLC细胞株H441,采用实时荧光定量PCR法检测不同组织及细胞中miR-363-3p相对表达量。将对数生长期的H441细胞分为对照组、miR-363-3p mimics组、miR-363-3p inhibitor组,分别使用Lipofectamine 2000转染试剂盒转染对照miRNA、miR-363-3p mimics、miR-363-3p inhibitor。转染30 h,采用Transwell法检测细胞迁移和侵袭能力(以迁移和侵袭细胞数表示),采用Western blotting法检测迁移标志物E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)及侵袭标志物基质金属蛋白酶9(MMP-9)、基质金属蛋白酶3(MMP-3)蛋白表达。结果发生和未发生淋巴结转移的NSCLC组织miR-363-3p相对表达量分别为1.091±0.153、0.128±0.336,二者比较P<0.01。A549、H441细胞miR-363-3p相对表达量分别为1.02±0.039、31.45±1.158,二者比较P<0.01。miR-363-3p inhibitor组、对照组、miR-363-3p mimics组迁移细胞数、侵袭细胞数及Vimentin、MMP-9、MMP-3蛋白相对表达量均依次升高,E-cadherin蛋白相对表达量依次降低,两组间比较P均<0.05。结论 miR-363-3p可能通过升高Vimentin、MMP-9、MMP-3蛋白表达及降低E-cadherin蛋白表达而促进NSCLC细胞的迁移和侵袭。Objective To observe the effects of miR-363-3p on migration and invasion of non-small-cell lung cancer(NSCLC).Methods Twenty-six cases of NSCLC tissues with lymph node metastasis and 21 case of NSCLC tissues with-out lymph node metastasis were collected.Human NSCLC cell lines A549 and H441 were cultured in vitro,and the expres-sion levels of miR-363-3p in different tissues and cells were detected by real-time fluorescence quantitative PCR.H441 cells in logarithmic growth phase were divided into the control group,miR-363-3p mimics group,and miR-363-3p inhibi-tor group,which were then transfected with the control miRNA,miR-363-3p mimics,and miR-363-3p inhibitor by Lipo-fectamine 2000 transfection kit,respectively.After 30 h of transfection,the cell migration and invasion abilities(ex-pressed by the number of migration and invasion cells)were detected by Transwell assay,and the expression levels of mi-gration markers E-cadherin(E-cadherin),Vimentin(Vimentin)and invasion markers matrix metalloproteinase 9(MMP-9),matrix metalloproteinase 3(MMP-3)protein by Western blotting.Results The expression levels of miR-363-3p in the NSCLC tissues with and without lymph node metastasis were 1.091±0.153 and 0.128±0.336,respectively,and the difference was statistically significant(both P<0.01).The expression levels of miR-363-3p in A549 and H441 cells were 1.02±0.039 and 31.45±1.158,and the difference was statistically significant(both P<0.01).In the miR-363-3p inhibi-tor group,control group,and miR-363-3p mimics group,the number of migrating cells,the number of invading cells and the relative expression levels of Vimentin,MMP-9,MMP-3 protein increased in sequence,while the expression level of Ecadherin protein decreased,with statistically significant difference(all P<0.05).Conclusion MiR-363-3p may promote the migration and invasion of NSCLC cells by increasing the expression of Vimentin,MMP-9,MMP-3 protein and reducing the expression of E-cadherin protein.

关 键 词：非小细胞肺癌 miR-363-3p 迁移 侵袭 E-CADHERIN VIMENTIN MMP-9 MMP-3 

分 类 号：R734.2[医药卫生—肿瘤]