miR-21靶向PDCD4对舌鳞癌细胞上皮间质转化及侵袭能力的影响  

Effects of miR-21 on epithelial mesenchymal transition and invasion ability of tongue squamous cell cells through targeting PDCD4

作　　者：丁文通 刘莹[1] 刘思瑶 姬红娇 潘爽 DING Wentong;LIU Ying;LIU Siyao;JI Hongjiao;PAN Shuang(Department of Endodontics,the First Affiliated Hospital of Harbin Medical University,Heilongjiang Harbin 150001,China)

机构地区：[1]哈尔滨医科大学附属第一医院牙体牙髓病科,黑龙江哈尔滨150001

出　　处：《现代肿瘤医学》2023年第24期4496-4503,共8页Journal of Modern Oncology

基　　金：国家自然科学基金(编号:81970924)。

摘　　要：目的:探究微小RNA-21(microRNA-21,miR-21)靶向程序性细胞死亡因子4(programmed cell death 4,PDCD4)对舌鳞癌细胞(tonguel squamous cell carcinoma,TSCC)上皮间质转化(epithelial-mesenchymal transition,EMT)及侵袭能力的影响。方法:qRT-PCR实验检测人口腔上皮细胞(human oral epithelial cell,HOEC)与舌鳞癌细胞系CAL-27中miR-21和PDCD4的表达水平。将miR-21 inhibitor阴性对照(NC inhibitor)、miR-21抑制剂(miR-21 inhibitor)转染至CAL-27细胞中,qRT-PCR实验检测miR-21的表达水平;采用CCK-8实验检测细胞增殖情况;采用划痕实验和Transwell迁移实验检测细胞迁移能力;采用Transwell侵袭实验检测细胞侵袭能力;采用免疫印迹法(Western blot)和qRT-PCR实验评估PDCD4、EMT标志物E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)及基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)、基质金属蛋白酶9(matrix metalloproteinase 9,MMP-9)在蛋白和mRNA水平的表达情况。双荧光素酶报告基因实验验证miR-21与PDCD4的靶向关系。结果:CAL-27细胞中miR-21的表达明显高于HOEC细胞(P<0.001),而PDCD4表达低于HOEC细胞(P<0.001)。与对照组相比,抑制miR-21后miR-21的表达水平明显降低(P<0.001),并能显著抑制细胞的增殖、迁移、侵袭能力(P<0.05);抑制miR-21能上调CAL-27细胞中PDCD4(P<0.05)和E-cadherin(P<0.05)的表达,而下调N-cadherin(P<0.05)、Vimentin(P<0.01)、MMP-2(P<0.01)、MMP-9(P<0.01)蛋白和mRNA水平。双荧光素酶报告基因实验证明miR-21与PDCD4的靶向关系。结论:抑制舌鳞癌细胞中miR-21的表达能抑制其增殖、侵袭和上皮间充质转化,这可能与靶向激活和促进PDCD4的表达有关。Objective:To investigate the effect of microRNA-21(miR-21)targeting programmed cell death 4(PDCD4)on epithelial-mesenchymal transition(EMT)and invasion ability of tongue squamous cell carcinoma(TSCC)cells.Methods:The expression levels of miR-21 and PDCD4 were detected by qRT-PCR experiment in human oral epithelial cells(HOEC)and tongue squamous cell carcinoma cell line CAL-27.The miR-21 inhibitor negative control(NC inhibitor)and miR-21 inhibitor were transfected into CAL-27 cells,and the expression level of miR-21 was detected by qRT-PCR experiment.Cell proliferation was detected by CCK-8 experiment,and cell migration ability was detected by scratch experiment and Transwell migration experiment.Cell invasion ability was detected by Transwell invasion experiment.Western blot and qRT-PCR experiment were used to evaluate the expression levels of PDCD4,EMT markers E-cadherin,N-cadherin,Vimentin,and matrix metalloproteinase 2(MMP-2),matrix metalloproteinase 9(MMP-9)at the protein and mRNA levels.The dual luciferase reporter gene experiment verified the targeting relationship between miR-21 and PDCD4.Results:The expression of miR-21 was significantly higher in CAL-27 cells than in HOEC cells(P<0.001),while the expression of PDCD4 was lower than in HOEC cells(P<0.001).Compared with the control group,the expression level of miR-21 was significantly reduced after inhibiting miR-21(P<0.001),which could significantly inhibit the proliferation,migration,and invasion abilities of cells(P<0.05).Inhibiting miR-21 could upregulate the expression of PDCD4(P<0.05)and E-cadherin(P<0.05)in CAL-27 cells,while downregulating the protein and mRNA levels of N-cadherin(P<0.05),Vimentin(P<0.01),MMP-2(P<0.01),and MMP-9(P<0.01).The dual luciferase reporter gene experiment confirmed the targeting relationship between miR-21 and PDCD4.Conclusion:Inhibiting the expression of miR-21 in tongue squamous cell carcinoma cells can inhibit their proliferation,invasion,and epithelial-mesenchymal transition,which may be related to the targeting activa

关 键 词：MIR-21 PDCD4 舌鳞癌细胞 EMT 迁移 侵袭 

分 类 号：R739.86[医药卫生—肿瘤]