丙泊酚介导miR-137/FGF9通路对人黑色素瘤细胞增殖、侵袭及迁移能力的影响

Effect of propofol on proliferation,invasion and migration of human melanoma cell via miR-137/FGF9 pathway

目的探讨丙泊酚介导miR-137/FGF9通路对人黑色素瘤细胞A375增殖、侵袭及迁移能力的影响。方法体外培养A375细胞,采用MTT法确定丙泊酚对A375细胞的半数抑制浓度和半数抑制时间,lipofectamine法分别将miR-137 mimics和miR-137 inhibitors转染到A375细胞。将A375细胞分为对照组、丙泊酣组、miR-137 mimics组、丙泊酣+miR-137 mimics组、miR-137 inhibitors组和丙泊酚+miR-137 inhibitors组。选用最佳浓度和时间分别进行相应处理后用实时荧光定量PCR(qRT-PCR)检测miR-137和FGF9 mRNA表达,同时Transwell法检测细胞体外侵袭和迁移能力。结果随着丙泊酚浓度增加,A375细胞增殖率降低,选80μmol/L作为半数抑制浓度;随着丙泊酚作用时间的增长,A375细胞增殖率降低,选48 h作为半数抑制时间。与对照组比较,丙泊酚可以促进miR-137 mRNA表达,抑制FGF9 mRNA表达,抑制A375细胞侵袭和迁移能力(P<0.05);miR-137 mimics可以促进miR-137 mRNA表达,抑制FGF9 mRNA表达,抑制A375细胞侵袭和迁移能力,同时给予丙泊酚干预后,促进miR-137 mRNA表达、抑制FGF9 mRNA表达、抑制A375细胞侵袭和迁移能力的效果更显著(P<0.05);miR-137 inhibitors可以抑制miR-137 mRNA表达,促进FGF9 mRNA表达,促进A375细胞侵袭和迁移能力(P<0.05),同时给予丙泊酚干预后,抑制miR-137 mRNA表达、促进FGF9 mKNA表达、促进A375细胞侵袭和迁移能力的效果受到一定的抑制(P<0.05)。结论丙泊酚对人黑色素瘤细胞A375增殖、侵袭及迁移能力具有明显的抑制作用,其机制可能与丙泊酚抑制miR-137/FGF9通路的激活有关。

Objective To investigate the effect of propofol on proliferation,invasion and migration of human melanoma cell line A375 via miR-137/fibroblast growth factor 9(FGF9)pathway.Methods A375 cells were cultured in vitro.The half inhibitory concentration and half inhibitory time of propofol on A375 cells were determined by methyl thiazolyl tetrazolium(MTT)method.miR-137 mimics and miR-137 inhibitors were transfected into A375 cells by lipofectamine method.A375 cells were divided into control group,propofol group,miR-137 mimics group,propofol+miR-137 mimics group,miR-137 inhibitors group and propofol+miR-137 inhibitors group.After treated with the optimal time and concentration,the mRNA expression of miR-137 and FGF9 was detected by real time fluorescence quantitative reverse transcription polymerase chain reaction(qRT-PCR),and the invasion and migration ability of cells in vitro was detected by transwell method.Results With the increase of propofol concentration,the proliferation rate of A375 cells was gradually decreased,and 80μmol/L was selected as the half inhibition concentration.With the increase of propofol action time,the proliferation rate of A375 cells was gradually decreased,and 48 hours was selected as the half inhibition time.Compared with the control group,propofol could promote the expression of miR-137 mRNA,inhibit the expression of FGF9 mRNA,and inhibit the invasion and migration of A375 cells(P<0.05).miR-137 mimics could promote the expression of miR-137 mRNA,inhibit the expression of FGF9 mRNA,and inhibit the invasion and migration of A375 cells.At the same time,after propofol intervention,the effect of promoting the expression of miR-137 mRNA,inhibiting the expression of FGF9 mRNA and inhibiting the invasion and migration of A375 cells was more significant(P<0.05);miR-137 inhibitors could inhibit the expression of miR-137 mRNA,promote the expression of FGF9 mRNA,and promote the invasion and migration of A375 cells(P<0.05).At the same time,after propofol intervention,the effects of inhibiting the expression of miR-137 mRNA,promoting the expression of FGF9 mRNA and promoting the invasion and migration of A375 cells were inhibited(P<0.05).Conclusions Propofol can inhibit the proliferation,invasion and migration of human melanoma cell line A375.The mechanism may be related to the inhibition of miR-137/FGF9 pathway activation by propofol.