利多卡因通过调控lncRNA TTN-AS1/miR-524-5p表达对骨肉瘤细胞增殖、迁移和侵袭的影响

目的:研究利多卡因对骨肉瘤细胞MG-63增殖、迁移和侵袭的影响，并探索其作用机制。方法:使用1 mmol/L、5 mmol/L、10 mmol/L浓度利多卡因处理MG-63细胞，MTT法检测细胞增殖，Transwell小室法检测细胞迁移和侵袭，Western blot检测细胞周期蛋白D1（CyclinD1）、p21、基质金属蛋白酶-2（MMP-2）、基质金属蛋白酶-9（MMP-9）蛋白表达，qRT-PCR检测长链非编码RNA TTN-AS1（TTN-AS1）和微小RNA-524-5p（miR-524-5p）表达，starBase软件结合双荧光素酶报告实验分析TTN-AS1与miR-524-5p的靶向关系。MG-63细胞中转染si-TTN-AS1、miR-524-5p或pcDNA-TTN-AS1（并进行10 mmol/L利多卡因处理），观察细胞的增殖、迁移、侵袭。结果:不同浓度利多卡因明显提高细胞增殖抑制率和p21蛋白表达量（P＜0.05），显著减少迁移细胞数、侵袭细胞数和CyclinD1、MMP-2、MMP-9蛋白表达量（P＜0.05），均呈剂量依赖性。TTN-AS1可靶向调控miR-524-5p表达。抑制TTN-AS1表达与miR-524-5p过表达显著增加MG-63细胞的增殖抑制率和p21蛋白表达量（P＜0.05），明显降低迁移细胞数、侵袭细胞数和CyclinD1、MMP-2、MMP-9蛋白表达量（P＜0.05）。TTN-AS1过表达逆转了利多卡因对MG-63细胞增殖、迁移、侵袭和CyclinD1、MMP-2、MMP-9蛋白表达的抑制作用，以及对miR-524-5p、p21蛋白表达的促进作用。结论:利多卡因通过调控lncRNA TTN-AS1/miR-524-5p表达抑制骨肉瘤MG-63细胞增殖、迁移和侵袭。

Effects of lidocaine on proliferation,migration and invasion of osteosarcoma cells by regulating lncRNA TTN-AS1/miR-524-5p expression

Objective:To study the effects of lidocaine on the proliferation,migration and invasion of osteosarcoma cell line MG-63,and to explore its mechanism.Methods:MG-63 cells were treated with 1 mmol/L,5 mmol/L,10 mmol/L lidocaine.Cell proliferation was detected by MTT assay.Cell migration and invasion were determined by Transwell chamber assay.CyclinD1,p21,MMP-2 and MMP-9 protein expression were detected by Western blot.qRT-PCR was used to detect TTN-AS1 and miR-524-5p expression,and starBase software combined with dual luciferase reporter assay was applied to analyze the targeting relationship between TTN-AS1 and miR-524-5p.The MG-63 cells were transfected with si-TTN-AS1,miR-524-5p or pcDNA-TTN-AS1 (treated with 10 mmol/L lidocaine) to observe cell proliferation,migration and invasion.Results:Different concentrations of lidocaine significantly increased the cell proliferation inhibition rate and p21 protein expression (P＜0.05),and evidently decreased the number of migrated cells,the number of invasive cells and the expression of CyclinD1,MMP-2 and MMP-9 (P＜0.05),both in a dose-dependent manner.TTN-AS1 targeted regulation of miR-524-5p expression.Inhibition of TTN-AS1 expression greatly improved the proliferation inhibition rate and p21 protein expression of MG-63 cells (P＜0.05),while obviously reduced the number of migrated cells,the number of invasive cells and CyclinD1,MMP-2,MMP-9 protein expression (P＜0.05),which was the same as the overexpression of miR-524-5p.TTN-AS1 overexpression reversed the inhibitory effect of lidocaine on MG-63 cell proliferation,migration,invasion and CyclinD1,MMP-2,MMP-9 protein expression,and reversed the promotion effect of miR-524-5p and p21 protein expression.Conclusion:Lidocaine inhibits the proliferation,migration and invasion of osteosarcoma MG-63 cells by regulating lncRNA TTN-AS1/miR-524-5p expression.