lncRNA FOXD2-AS1靶向miR-1299抑制口腔鳞癌细胞增殖和诱导凋亡的体外研究

In vitro study of lncRNA FOXD2AS1 targeting miR1299 in inhibiting the proliferation of oral squamous cell carcinoma cells and inducing apoptosis

目的探讨lncRNAFOXD2-AS1靶向miR-1299抑制口腔鳞癌细胞增殖和诱导凋亡的体外研究。方法实验分为si-NC组、si-FOXD2-AS1组、pcDNA-NC组、pcDNA-FOXD2-AS1组、miR-NC组、miR-1299组、anti-miR-NC+si-FOXD2-AS1组、anti-miR-1299+si-FOXD2-AS1组。实时荧光定量PCR(qPCR)检测lncRNAFOXD2-AS1和miR-1299的表达水平。细胞计数试剂盒8(CCK-8)检测细胞增殖能力。流式细胞术检测细胞凋亡。双荧光素酶报告实验检测lncRNAFOXD2-AS1与miR-1299的靶向作用。Western blot法检测CyclinD1、Cleaved-caspase-3、β-连环蛋白(β-catenin)蛋白表达。结果口腔鳞癌细胞CAL27、SCC-090、HSC-3中lncRNAFOXD2-AS1表达水平显著升高,miR-1299表达水平显著降低(P<0.05)。lncRNAFOXD2-AS1低表达或miR-1299高表达均可抑制口腔鳞癌细胞增殖,促进细胞凋亡。lncRNAFOXD2-AS1靶向调控miR-1299,低表达miR-1299可以逆转lncRNAFOXD2-AS1低表达对口腔鳞癌细胞增殖和凋亡的影响。lncRNAFOXD2-AS1低表达抑制β-catenin蛋白表达,而低表达miR-1299逆转了lncRNAFOXD2-AS1低表达对β-catenin蛋白表达的抑制作用。结论干扰lncRNAFOXD2-AS1表达可能通过上调miR-1299抑制Wnt/β-catenin信号通路抑制口腔鳞癌细胞增殖,促进细胞凋亡。

Objective To investigate the effects of lncRNAFOXD2-AS1 targeting miR-1299 in inhibiting the proliferation of oral squamous cell carcinoma(OSCC)cells and inducing apoptosis in vitro.Methods The experiment was divided into si-NC group,si-FOXD2-AS1 group,pcDNA-NC group,pcDNA-FOXD2-AS1 group,miR-NC group,miR-1299 group,anti-miR-NC+si-FOXD2-AS1 group,and anti-miR-1299+si-FOXD2-AS1 group.Real-time fluorescent quantitative PCR(RT-qPCR)was used to detect the expression levels of lncRNAFOXD2-AS1 and miR-1299.Cell counting kit 8(CCK 8)was used to detect cell proliferation ability,and flow cytometry was used to detect cell apoptosis.The dual-luciferase reporter assay system was used to detect the targeting effect of lncRNAFOXD2-AS1 on miR-1299.Western blot method was used to detect the protein expression of CyclinD1,Cleaved-caspase-3,andβ-catenin.Results The expression levels of lncRNAFOXD2-AS1 in OSCC cells CAL27,SCC-090 and HSC-3 were significantly increased,while the expression levels of miR-1299 were significantly decreased(P<0.05).Low expression of lncRNAFOXD2-AS1 or high expression of miR-1299 could inhibit the proliferation of OSCC cells and promote cell apoptosis.LncRNAFOXD2-AS1 targeted and regulated miR-1299,and low expression of miR-1299 could reverse the effects of low expression of lncRNAFOXD2-AS1 on the proliferation and apoptosis of OSCC cells.Low expression of lncRNAFOXD2-AS1 inhibitedβcatenin protein expression,while low expression of miR-1299 reversed the inhibitory effect of low expression of lncRNAFOXD2-AS1 onβcatenin protein expression.Conclusion Interfering with the expression of lncRNAFOXD2 AS1 may inhibit the proliferation of OSCC cells and promote apoptosis by up-regulating miR-1299 and inhibiting the Wnt/β-catenin signaling pathway.