利多卡因调控微小RNA-195对骨肉瘤MG63细胞增殖、凋亡及化疗敏感性的影响

Effect of lidocaine of proliferation,apoptosis and chemosensitivity of osteosarcoma MG63 cells by regulating miR-195

目的探讨利多卡因(lidocaine)对骨肉瘤MG63细胞增殖、凋亡及化疗敏感性的影响及其可能的作用机制。方法研究于2018年5―12月,体外培养骨肉瘤MG63细胞,不同浓度的利多卡因干预MG63细胞,采用甲基噻唑基四唑(MTT)法检测MG63细胞存活率及顺铂半抑制浓度(IC50值);流式细胞术检测细胞凋亡率;实时荧光定量聚合酶链反应(qRT-PCR)检测微小RNA-195(miR-195)、多药耐药相关蛋白1(MRP1)的表达水平;观察miR-195过表达或抑制miR-195的表达后MG63细胞增殖、凋亡及IC50值变化情况;蛋白质印迹法(Westernblotting)检测增殖、凋亡相关蛋白表达。结果利多卡因对MG63细胞增殖具有一定抑制作用;与0 mmol/L组相比,利多卡因2 mmol/L组、4 mmol/L组细胞凋亡率[(18.35±1.85)%、(30.25±3.03)%比(8.22±0.82)%]、活化的半胱氨酸天冬氨酸蛋白酶3(Cleaved-caspase-3)[(0.65±0.06)、(1.00±0.10)比(0.15±0.02)]与miR-195的表达水平增加[(1.86±0.18)、(2.25±0.22)比(1.00±0.10)],MRP1 mRNA表达水平[(0.72±0.08)、(0.42±0.05)比(1.00±0.11)]、顺铂IC50值降低[(6.59±0.71)mg/L、(2.16±0.23)mg/L比(9.05±0.92)mg/L],差异有统计学意义(P<0.05),MG63细胞中miR-195过表达具有相似作用;抑制miR-195的表达可逆转利多卡因对MG63细胞增殖、凋亡及化疗敏感性的作用。结论利多卡因可通过miR-195抑制骨肉瘤MG63细胞增殖及诱导细胞凋亡,并可增强其对顺铂化疗敏感性。

Objective To investigate the effect of lidocaine on proliferation,apoptosis and chemosensitivity of osteosarcoma MG63 cells and its possible mechanism.Methods The study started May 2018 to December 2018,osteosarcoma MG63 cells were cultured in vitro,and MG63 cells were treated with different concentrations of lidocaine.MTT was used to detect the survival rate of MG63 cells and the semi-inhibitory concentration of cisplatin(IC50 value).Flow cytometry was used to detect the apoptosis rate.Real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression levels of microRNA-195(miR-195)and multidrug resistance-associated protein 1(MRP1).The proliferation,apoptosis and IC50 value of MG63 cells were observed after overexpression of miR-195 or inhibition of miR-195 expression.The proliferation and apoptosis-related protein expression were detected by Western blot.Results Lidocaine had a certain inhibitory effect on the proliferation of MG63 cells.Compared with the 0 mmol/L group,the apoptosis rate of lidocaine[(18.35±1.85)%,(30.25±3.03)%vs.(8.22±0.82)%],Cleaved-caspase-3[(0.65±0.06),(1.00±0.10)vs.(0.15±0.02)]and the expression levels of miR-195[(1.86±0.18),(2.25±0.22)vs.(1.00±0.10)]in 2 mmol/L,4 mmol/L groups were increased,MRP1 mRNA expression levels[(0.72±0.08),(0.42±0.05)vs.(1.00±0.11)],cisplatin IC50 value[(6.59±0.71)mg/L,(2.16±0.23)mg/L vs.(9.05±0.92)mg/L]were decreased,the difference was statistically significant(P<0.05),and miR-195 overexpression in MG63 cells had a similar effect.Inhibition of miR-195 expression reversed the effect of lidocaine on proliferation,apoptosis and chemosensitivity of MG63 cells.Conclusion Lidocaine can inhibit the proliferation of osteosarcoma MG63 cells and induce apoptosis by regulating miR-195,and enhance its sensitivity to cisplatin chemotherapy.