LINC00662通过调控miR-215-3p/EIF4H轴影响骨肉瘤细胞的增殖和凋亡

LINC00662 affects the proliferation and apoptosis of osteosarcoma cells by regulating miR-215-3P/EIF4H axis

目的:探讨长基因间非蛋白质编码RNA 662(LINC00662)对骨肉瘤细胞的增殖和凋亡的影响及可能机制。方法:实时荧光定量聚合酶链反应(RT-qPCR)和Western blotting检测人成骨细胞株hFOB 1.19和骨肉瘤细胞中LINC00662、miR-215-3p和真核生物翻译起始因子4H(EIF4H)表达水平。将LINC00662小干扰RNA(si-LINC00662)、miR-215-3p模拟物、EIF4H小干扰RNA(si-EIF4H)分别转染Saos2细胞,细胞计数试剂盒(CCK-8)检测细胞活力,流式细胞术检测细胞凋亡。双荧光素酶报告实验、RT-qPCR、Western blotting确定LINC00662与miR-215-3p、miR-215-3p与EIF4H之间的靶向关系。结果:与hFOB 1.19比较,骨肉瘤细胞中LINC00662和EIF4H表达显著升高,miR-215-3p表达显著降低(P<0.05)。抑制LINC00662表达或过表达miR-215-3p或抑制EIF4H表达后,Saos2细胞存活率均显著降低,凋亡率均显著升高(P<0.05)。LINC00662靶向负性调控miR-215-3p表达,miR-215-3p靶向负性调控EIF4H表达(P<0.05)。抑制miR-215-3p表达可逆转抑制miR-215-3p对Saos2细胞存活和凋亡的影响(P<0.05)。过表达EIF4H可逆转抑制miR-215-3p对Saos2细胞存活和凋亡的影响(P<0.05)。结论:干扰LINC00662表达可抑制骨肉瘤细胞增殖,促进细胞凋亡,其机制与靶向调控miR-215-3p/EIF4H轴有关。

Objective:To investigate the effect of long intergene non-protein coding RNA 662(LINC00662)on the proliferation and apoptosis of osteosarcoma cells,and to explore its possible mechanism further.Methods:Real-time fluorescence quantitative PCR(RT-qPCR)and Western blotting were used to detect the expression level of LINC00662,miR-215-3p and Eukaryotic translation initiation factor 4H(EIF4H)in human osteoblast cell line hFOB 1.19 and osteosarcoma cells.LINC00662 small interfering RNA(si-LINC00662),miR-215-3p mimic,EIF4H small interfering RNA(si-EIF4H)were transfected into Saos2 cells,respectively.Cell counting kit(CCK-8)detected cell viability and flow cytometry detected cell apoptosis.The dual luciferase report experiment,RT-qPCR and Western blotting were applied to confirm the targeting relationship between LINC00662 and miR-215-3p,miR-215-3p and EIF4H.Results:Compared with hFOB 1.19,the expression of LINC00662 and EIF4H in osteosarcoma cells were significantly increased,while the expression of miR-215-3p was significantly reduced(P<0.05).After inhibiting the expression of LINC00662 or overexpressing miR-215-3p or inhibiting the expression of EIF4H,the survival rate of Saos2 cells was significantly reduced,and the apoptosis rate was significantly increased(P<0.05).LINC00662 targets and negatively regulates miR-215-3p expression,and miR-215-3p targets and negatively regulates EIF4H expression(P<0.05).Inhibiting the expression of miR-215-3p could reverse the effect of inhibiting miR-215-3p on the survival and apoptosis of Saos2 cells(P<0.05).Overexpression of EIF4H could reverse the effect of inhibiting miR-215-3p on the survival and apoptosis of Saos2 cells(P<0.05).Conclusion:Interfering with the expression of LINC00662 could inhibit osteosarcoma cells proliferation and promote cell apoptosis,and its mechanism was related to the targeted regulation of miR-215-3p/EIF4H axis.