山东医药2024，Vol.64Issue(18)：26-30,5.DOI:10.3969/j.issn.1002-266X.2024.18.006
lncRNA NEAT1降表达人喉鳞状细胞癌细胞增殖凋亡变化及与miR-214靶向关系
Changes in proliferation and apoptosis of human laryngeal squamous cell carcinoma cells with down-expression of LncRNA NEAT1 and its targeted relationship with miR-214
摘要
Abstract
Objective To observe the effects of long non-coding RNA(lncRNA)nuclear-enriched abundant tran-script 1(NEAT1)on the proliferation and apoptosis of human laryngeal squamous cell carcinoma(LSCC)cells,as well as its targeted relationship with microRNA-214(miR-214).Methods Real-time fluorescence quantitative PCR(RT-qP-CR)was used to detect lncRNA NEAT1 and miR-214 in immortalized human epidermal cells Hacat and human LSCC cell lines(FD-LSC-1,Hep-2,and TU177),and then TU177 cells were selected as the experimental cells.TU177 cells in the logarithmic growth phase were divided into groups A and B,which were transfected with si-lncRNA NEAT1(knockdown of lncRNA NEAT1 expression)and si-NC(random meaningless sequence),respectively.At 24 h,cell proliferation was ob-served using CCK-8 assay,colony formation experiment was conducted to observe clone formation,flow cytometry was used to analyze cell cycle distribution and apoptosis rate,while RT-qPCR was performed to detect lncRNA NEAT1 and miR-214 expression in both groups.TU177 cells in the logarithmic growth phase were divided into groups Ⅰ,Ⅱ,Ⅲ and Ⅳ,which were transfected with WT-lncRNA NEAT1 and miR-214 mimic,WT-lncRNA NEAT1 and miR-NC,MUT-lncRNA NEAT1 and miR-214 mimic,and MUT-lncRNA NEAT1 and miR-NC,respectively.Th targeted relationship between ln-cRNA NEAT1 and miR-214 was validated by dual luciferase reporter gene assay.Results Compared with the group B,the OD values of TU177 cells were lower at 24,48,and 72 h of cultivation;at 14 d of cultivation,the number of clone for-mation in TU177 cells was smaller;the proportion of cells in the G0/G1 phase was higher,the proportion of cells in the S phase was lower,and the apoptosis rate was higher in the group A(all P<0.05).Compared with the group B,at 24 h of trans-fection,the relative expression of lncRNA NEAT1 was lower and the relative expression of miR-214 in TU177 cells was higher in the group A(both P<0.05).At 48 h of cultivation,the luciferase activity of TU177 cells in the groups Ⅰ,Ⅱ,Ⅲand Ⅳ was 0.63±0.08,0.99±0.01,1.02±0.02,and 0.98±0.03,respectively.Compared with the group Ⅰ,groupⅡ had lower cell luciferase activity(P<0.05).Conclusions Knockdown of lncRNA NEAT1 expression can inhibit the proliferation and cloning of TU177 cells,block the cell cycle in G0/G1 phase,and promote the apoptosis.LncRNA NEAT1 is targetedly associated with miR-214 in TU177 cells.LncRNA NEAT1 may inhibit the proliferation and cloning of TU177 cells,block the cell cycle in G0/G1 phase,and promote the apoptosis by targetedly binding with miR-214.关键词
长链非编码RNA/长链非编码RNA 核富集转录体1/微小RNA-214/喉鳞状细胞癌/细胞增殖/细胞凋亡/细胞克隆/细胞周期Key words
long non-coding RNA/long non-coding RNA nuclear-enriched abundant transcript 1/microRNA-214/laryngeal squamous cell carcinoma/cell proliferation/apoptosis/cell cloning/cell cycle分类
医药卫生引用本文复制引用
罗德艳,冯俊,彭涛,唐一萍,杨久梅..lncRNA NEAT1降表达人喉鳞状细胞癌细胞增殖凋亡变化及与miR-214靶向关系[J].山东医药,2024,64(18):26-30,5.基金项目
四川省科技厅重点研发项目(2018FZ0116) （2018FZ0116）
四川省教育厅自然科学重点项目(18ZA203) （18ZA203）
南充市科技局应用技术研究与开发基金项目(16YFZJ0021). （16YFZJ0021）
