瑞芬太尼通过调控lncRNA SUMO1P3表达抑制肾癌细胞786-O的增殖、迁移和侵袭

Remifentanil inhibits proliferation,migration and invasion of renal cancer cells 786-O by regulating the expression of lncRNA SUMO1P3

目的探讨瑞芬太尼是否通过lncRNA SUMO1P3影响肾癌细胞786-O增殖、迁移和侵袭。方法体外培养786-O细胞,分为对照组(正常培养48 h)和不同浓度瑞芬太尼组(20,40,80 nmol/L瑞芬太尼的培养基培养48 h),细胞计数试剂盒-8(CCK-8)检测细胞活性,Transwell实验检测细胞迁移和侵袭,实时荧光定量PCR(RT-qPCR)检测细胞中小泛素样修饰蛋白1假基因3(SUMO1P3)表达,蛋白质印迹法(Western blot)检测细胞中细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶(MMP)-2和MMP-9蛋白表达。786-O细胞分别转染SUMO1P3小干扰RNA(si-SUMO1P3组)和小干扰RNA阴性对照序列(si-NC组)后,CCK-8、Transwell及Western blot法分别观察沉默SUMO1P3基因表达对786-O细胞活性、迁移和侵袭,及Cyclin D1、MMP-2和MMP-9蛋白表达的影响。786-O细胞分别转染SUMO1P3过表达载体(pcDNA-SUMO1P3)和空载体(pcDNA),然后再用80 nmol/L瑞芬太尼干预(依次记为80 nmol/L瑞芬太尼+pcDNA-SUMO1P3组、80 nmol/L瑞芬太尼+pcDNA组)后,CCK-8、Transwell及Western blot法分别观察细胞活性、迁移和侵袭,及Cyclin D1、MMP-2和MMP-9蛋白表达。结果与0 nmol/L组比较,20,40,80 nmol/L瑞芬太尼组786-O细胞活性、迁移数和侵袭数及细胞中Cyclin D1、MMP-2和MMP-9蛋白表达均降低(P<0.05),SUMO1P3基因表达降低(P<0.05),且不同浓度瑞芬太尼组间各检测指标比较差异均具有统计学意义(P<0.05)。与si-NC组比较,si-SUMO1P3组786-O细胞活性、迁移数和侵袭数,及细胞中Cyclin D1、MMP-2和MMP-9蛋白表达均降低,差异均具有统计学意义(P<0.05)。与80 nmol/L瑞芬太尼+pcDNA组比较,80 nmol/L瑞芬太尼+pcDNA-SUMO1P3组786-O细胞活性、迁移数和侵袭细胞数,及细胞中Cyclin D1、MMP-2和MMP-9蛋白表达均升高,差异均具有统计学意义(P<0.05)。结论瑞芬太尼可通过抑制SUMO1P3的表达阻碍肾癌786-O细胞增殖、迁移和侵袭。

Objective To explore whether remifentanil affects the proliferation,migration and invasion of renal cancer cell 786-O through lncRNA SUMO1P3.Methods 786-O cells were divided into control group and different concentrations of remifentanil groups,and cultured in vitro in a medium containing 0,20,40,80 nmol/L remifentanil,respectively.And then CCK-8 was used to detect the cell viability,Transwell was used to detect the cell migration and invasion,RT-qPCR was used to detect the expression level of SUMO1P3 in cells,and Western blot was used to detect the protein expression of Cyclin D1,MMP-2 and MMP-9.After 786-O cells were transfected with SUMO1P3 small interfering RNA(si-SUMO1P3 group)or small interfering RNA negative control sequence(si-NC group),CCK-8,Transwell and Western blot were used to observe the activity,migration and invasion of 786-O cells and the protein expression of Cyclin D1,MMP-2 and MMP-9.After transfected with SUMO1P3 overexpression vector(pcDNA-SUMO1P3)or empty vector(pcDNA),786-O cells were treated with 80 nmol/L remifentanil,named as 80 nmol/L remifentanil+pcDNASUMO1P3 group and 80 nmol/L remifentanil+pcDNA group,and then CCK-8,Transwell and Western blot were used to observe the activity,migration and invasion of 786-O cells and the protein expression of Cyclin D1,MMP-2 and MMP-9.Results Compared with 0 nmol/L group,the viability,the migration and invasion of 786-O cells,and the protein expression of Cyclin D1,MMP-2 and MMP-9 in 20,40,80 nmol/L remifentanil groups were decreased(P<0.05),and the expression of SUMO1P3 was decreased(P<0.05),and there was significant difference in the above indicators between different concentration of remifentanil groups(P<0.05).Compared with si-NC group,the viability,the numbers of migration and invasion of 786-O cells,and the protein expression of Cyclin D1,MMP-2 and MMP-9 in si-SUMO1P3 group were decreased(all P<0.05).Compared with 80 nmol/L remifentanil+pcDNA group,the viability,the migration and invasion of 786-O cells,and the protein expression of Cyclin D1,MMP-2 and MMP-9 in 80 nmol/L remifentanil+pcDNA-SUMO1P3 group were increased(P<0.05).Conclusion Remifentanil may inhibit the proliferation,migration and invasion of renal carcinoma 786-O cells by inhibiting the expression of SUMO1P3.