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
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.
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