紫铆因通过调控lncRNA STXBP5-AS1/miR-892a轴对口腔鳞癌细胞的糖酵解、凋亡、迁移及侵袭的影响

Effects of Butein on Glycolysis,Apoptosis,Migration and Invasion of Oral Squamous Cell Carcinoma Cells by Regulating lncRNA STXBP5-AS1/miR-892a Axis

目的:探讨紫铆因对口腔鳞癌细胞(HSC-3)糖酵解、凋亡、迁移及侵袭的影响和机制。方法:不同浓度紫铆因处理HSC-3细胞,运用流式细胞术、Transwell实验检测细胞凋亡、迁移和侵袭。试剂盒检测葡糖糖消耗以及乳酸产生量。实时荧光定量PCR (RT-qPCR)检测长链非编码RNA (lncRNA) STXBP5反义RNA1 (STXBP5-AS1)和miR-892a表达量。双荧光素酶报告实验和RT-qPCR分析和验证STXBP5-AS1与miR-892a调控关系。转染STXBP5-AS1过表达质粒至HSC-3细胞,检测STXBP5-AS1过表达对细胞糖酵解、凋亡、迁移及侵袭的影响。转染STXBP5-AS1小干扰RNA至HSC-3细胞,检测抑制STXBP5-AS1表达对紫铆因处理的HSC-3细胞糖酵解、凋亡、迁移及侵袭的影响。结果:紫铆因处理后HSC-3细胞葡萄糖消耗量、乳酸产生量、迁移和侵袭细胞数显著降低,凋亡率显著升高,STXBP5-AS1表达显著升高,miR-892a表达显著降低(P<0.05),并表现出一定剂量依赖效应。miR-892a是STXBP5-AS1的靶基因。过表达STXBP5-AS1后HSC-3细胞葡萄糖消耗量、乳酸产生量、迁移和侵袭细胞数显著降低,凋亡率显著升高,miR-892a表达显著降低(P<0.05)。抑制STXBP5-AS1表达可降低紫铆因处理对HSC-3细胞糖酵解、凋亡、迁移及侵袭的影响(P<0.05)。结论:紫铆因可抑制HSC-3细胞的迁移侵袭和糖酵解代谢,促进细胞凋亡,其机制可能与调控lncRNA STXBP5-AS1/miR-892a轴有关。

Objective: To investigate the effect and mechanism of butein on glycolysis, apoptosis, migration and invasion of oral squamous cell carcinoma(HSC-3) cells. Methods:HSC-3 cells were treated with different concentrations of butein,and cell apoptosis,migration and invasion were detected by flow cytometry and Transwell assay. The kit detects glucose consumption and lactic acid production. The expressions of long-chain noncoding RNA(lncRNA) STXBP5 antisense RNA1(STXBP5-AS1) and miR-892 a were detected by real-time fluorescence quantitative PCR(RT-q PCR). The regulatory relationship between STXBP5-AS1 and miR-892 a was analyzed and verified by double Luciferase Report experiment and RTqPCR. STXBP5-AS1 overexpression plasmid was transfected into HSC-3 cells to detect the effects of STXBP5-AS1 overexpression on cell glycolysis,apoptosis,migration and invasion. Transfect STXBP5-AS1 small interfering RNA into HSC-3 cells,and detect the effects of inhibiting the expression of STXBP5-AS1 on glycolysis,apoptosis,migration and invasion of butein treated HSC-3 cells. Results:After butein treatment,the glucose consumption,lactic acid production,migration and invasion of HSC-3 cells decreased significantly,the apoptosis rate increased significantly,the expression of STXBP5-AS1 increased significantly,and the expression of miR-892 a decreased significantly(P<0.05). miR-892 a is the target gene of STXBP5-AS1. After overexpression of STXBP5-AS1,glucose consumption,lactate production,migration and invasion of HSC-3 cells decreased significantly,the apoptosis rate increased significantly,and the expression of miR-892 a decreased significantly(P<0.05). Inhibition of STXBP5-AS1 expression could reduce the effects of butein treatment on glycolysis,apoptosis, migration and invasion of HSC-3 cells(P<0.05). Conclusion: Butein can inhibit the migration, invasion,glycolysis and metabolism of HSC-3 cells and promote cell apoptosis. Its mechanism may be related to the regulation of lncRNA STXBP5-AS1/miR-892 a axis.