LncRNA NORAD靶向miR-34a对口腔鳞癌细胞MMP-2表达的机制研究  

Mechanisms of LncRNA NORAD-targeted miR-34a on MMP-2 expression in oral squamous cell carcinoma cells

作　　者：陈赫 崔怡 靳琳玉 岳磊[2] CHEN He;CUI Yi;JIN Lin-yu;YUE Lei(The Fourth Hospital of Hebei Medical University,Hebei Shijiazhuang 050000,China;The Second Hospital of Hebei Medical University,Hebei Shijiazhuang 050000,China)

机构地区：[1]河北医科大学第四医院,河北石家庄050000 [2]河北医科大学第二医院,河北石家庄050000

出　　处：《临床口腔医学杂志》2023年第8期451-455,共5页Journal of Clinical Stomatology

基　　金：河北省医学科学研究基金项目(编号:20201112);河北省重点科技研究计划项目(编号:20221255)。

摘　　要：目的:探究lncRNA NORAD靶向miR-34a对口腔鳞状细胞癌MMP-2表达的影响。方法:将SCC-9细胞随机分为control组(不转染任何质粒)、si-NORAD(SCC-9细胞转染si-NORAD质粒)及si-NC组(SCC-9细胞转染阴性对照质粒)、miR-34a组(SCC-9细胞转染miR-34a mimics质粒)、si-NORAD+miR-34a inhibitor组(SCC-9细胞同时转染si-NORAD质粒和miR-34a inhibitor质粒)。RT-PCR检测细胞中NORAD、miR-34a表达;CCK-8检测细胞增殖能力;Transwell检测细胞侵袭能力;蛋白质印迹(Western blotting)检测细胞中MMP-2蛋白表达;荧光素酶实验报告检测NOARD和miR-34a的靶向关系。结果:与人永生化口腔上皮细胞HIOEC相比,口腔鳞癌细胞株中NORAD表达均明显增加,miR-34a表达明显降低,且SCC-9细胞中NORAD表达和miR-34a表达变化最显著(P<0.05)。与control组相比,si-NORAD组和miR-34a组细胞增殖、侵袭,以及MMP-2蛋白表达均明显降低(P<0.05);control组和si-NC组、si-NORAD组和miR-34a组细胞增殖、侵袭,以及MMP-2蛋白表达相比无明显差异(P>0.05);与si-NORAD组相比,si-NORAD+miR-34a inhibitor组细胞增殖、侵袭,以及MMP-2蛋白表达均明显增加(P<0.05)。结论:敲减NORAD可靶向调控miR-34a,进而抑制口腔鳞状细胞癌细胞的增殖、侵袭和细胞中MMP-2表达。Objective:To investigate the effect of lncRNA NORAD targeting miR-34a on MMP-2 expression in oral squamous cell carcinoma.Methods:SCC-9 cells were randomly divided into control group(not transfected with any plasmid),si-NORAD(SCC-9 cells transfected with si-NORAD plasmid)and si-NC group(SCC-9 cells transfected with negative control plasmid),miR-34a group(SCC-9 cells transfected with miR-34a mimics plasmid),si-NORAD+miR-34a inhibitor group(SCC-9 cells were transfected with both si-NORAD plasmid and miR-34a inhibitor plasmid).RT-PCR was performed to detect the expression of NORAD,miR-34a.CCK-8 to detect cell proliferation ability,Transwell to detect cell invasion ability,Western blotting to detect MMP-2 protein expression in cells,luciferase assay to detect the targeting relationship between NOARD and miR-34a.Results:Compared with human immortalized oral epithelial cells HIOEC,NORAD expression was significantly increased and miR-34a expression was significantly decreased in both oral squamous carcinoma cell lines,and the most significant changes in NORAD expression and miR-34a expression were observed in SCC-9 cells(P<0.05).Compared with the control group,cell proliferation,invasion,and MMP-2 protein expression were significantly decreased in the si-NORAD and miR-34a groups(P<0.05).There was no significant difference in cell proliferation,invasion,and MMP-2 protein expression in the control group compared with the si-NC group,si-NORAD group,and miR-34a group(P>0.05).Compared with the si-NORAD group cell proliferation,invasion,and MMP-2 protein expression were significantly increased in the si-NORAD+miR-34a inhibitor group(P<0.05).Conclusion:Knockdown of NORAD can target and regulate miR-34a,which in turn inhibits proliferation,invasion and MMP-2 expression in oral squamous cell carcinoma cells.

关 键 词：LncRNA NORAD MIR-34A 口腔鳞状细胞癌 基质金属蛋白酶2(MMP-2) 

分 类 号：R739.8[医药卫生—肿瘤]