lncRNA FOXD2-AS1靶向miR-3194-3p对口腔鳞癌细胞增殖及凋亡影响的体外研究

In vitro Study on the Effect of lncRNA FOXD2-AS1 Targeting MiR-3194-3p on the Proliferation and Apoptosis of Oral Squamous Cell Carcinoma

目的:探讨lncRNA FOXD2-AS1对口腔鳞癌细胞增殖及凋亡的影响及分子机制。方法:实时荧光定量PCR(RT-qPCR)检测口腔鳞癌细胞系中lncRNA FOXD2-AS1和miR-3194-3p的表达水平;将FOXD2-AS1干扰表达载体、miR-3194-3p模拟物、miR-3194-3p抑制剂+FOXD2-AS1干扰表达载体转染至CAL27细胞中,Western blot法检测蛋白表达;MTT检测细胞存活率;流式细胞术检测细胞凋亡;双荧光素酶报告实验检测lncRNA FOXD2-AS1和miR-3194-3p的靶向关系。结果:口腔鳞癌细胞系中lncRNA FOXD2-AS1表达水平升高,miR-3194-3p表达水平降低(P<0.01)。lncRNA FOXD2-AS1低表达或miR-3194-3p高表达,口腔鳞癌CAL27细胞中细胞周期蛋白D1(Cyclin D1)表达水平和细胞存活率降低,裂解的半胱氨酸天冬氨酸蛋白酶-3(cleaved-caspase-3)表达水平和细胞凋亡率升高(P<0.01);且lncRNA FOXD2-AS1低表达降低了β-连环蛋白(β-catenin)蛋白表达。lncRNA FOXD2-AS1靶向调控miR-3194-3p,低表达miR-3194-3p逆转了FOXD2-AS1低表达对口腔鳞癌细胞CAL27增殖和凋亡及Wnt/β-catenin信号通路的影响。结论:下调lncRNA FOXD2-AS1表达可能通过上调miR-3194-3p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 explore the effect and molecular mechanism of lncRNA FOXD2-AS1 in the proliferation and apoptosis of oral squamous carcinoma cells.Methods: Real-time fluorescent quantitative PCR(RT-qPCR) was used to detect the expression levels of lncRNA FOXD2-AS1 and miR-3194-3p in oral squamous cell carcinoma cell lines;the FOXD2-AS1 interference expression vector,miR-3194-3p mimic,miR-3194-3p inhibitor and FOXD2-AS1 interference expression vector were transfected into CAL27 cells,and the protein expression was detected by Western blot;MTT was used to detect the cell viability;flow cytometry was used to detect the cell apoptosis;dual luciferase reporter experiment was applied to detect the targeting relationship between lncRNA FOXD2-AS1 and miR-3194-3p.Results: The expression of lncRNA FOXD2-AS1 in oral squamous cell carcinoma cell lines was increased,and the expression of miR-3194-3p was decreased(P<0.01).With low expression of lncRNA FOXD2-AS1 or high expression of miR-3194-3 p,Cyclin D1 expression and cell survival rate in oral squamous cell carcinoma CAL27 cells were decreased,and leaved-caspase-3 expression and cell apoptosis rate were increased(P<0.01);and the low expression of lncRNA FOXD2-AS1 reduced the expression of β-catenin protein.LncRNA FOXD2-AS1 targeted and regulated miR-3194-3 p.The low expression of miR-3194-3 p reversed the effects of low FOXD2-AS1 expression on the proliferation and apoptosis of oral squamous cell carcinoma cells CAL27 and Wnt/β-catenin signaling pathway.Conclusion: Down-regulation of lncRNA FOXD2-AS1 expression may inhibit the proliferation of oral squamous carcinoma cells and promote the apoptosis by up-regulating miR-3194-3 p to inhibit Wnt/β-catenin signaling pathway.