IncRNA 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.

Objective To observe the effects of long non-coding RNA (IncRNA) nuclear-enriched abundant transcript 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 IncRNA NEATI 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-IncRNA NEATI (knockdown of IncRNA NEATI expression) and si-NC (random meaningless sequence), respectively. At 24 h, cell proliferation was observed 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 IncRNA NEATI 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-IncRNA NEAT1 and miR-214 mimic, WT-IncRNA NEATI and miR-NC, MUT-IncRNA NEAT1 and miR-214 mimic, and MUT-IncRNA NEAT1 and miR-NC, respectively. Th targeted relationship between In-CRNA NEATI 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 formation in TU177 cells was smaller; the proportion of cells in the Go/G, 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 transfection, the relative expression of IncRNA NEATI 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 TU 177 cells in the groups I, II, III and 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, group II had lower cell luciferase activity (P<0.05). Conclusions Knockdown of IncRNA NEATI expression can inhibit the proliferation and cloning of TU177 cells, block the cell cycle in Go/G, phase, and promote ote the apoptosis. LncRNA NEATI is targetedly associated with miR-214 in TU177 cells. LncRNA NEATI may inhibit the proliferation and cloning of TU 177 cells, block the cell cycle in Go/G, phase, and promote the apoptosis by targetedly binding with miR-214. [ABSTRACT FROM AUTHOR]