顺阿曲库铵通过miR-526b-3p/FMNL3通路抑制胃癌AGS细胞增殖、迁移和侵袭

Cisatracurium regulates proliferation,migration and invasion of gastric cancer AGS cells via miR-526b-3p/FMNL3 pathway

目的:探讨顺阿曲库铵对胃癌AGS细胞增殖、迁移和侵袭的影响及机制。方法:选取2019年1月至2022年1月胃癌手术切除组织及癌旁正常胃黏膜组织30例。实验设置对照组(正常培养)、顺阿曲库铵低(10 mmol/L)、中(20 mmol/L)、高(30 mmol/L)浓度组、miR-NC组、miR-526b-3p组、anti-miR-NC组、anti-miR-526b-3p组、Cis+anti-miR-NC组、Cis+anti-miR-526b-3p组。Cis+anti-miR-NC组、Cis+anti-miR-526b-3p组分别转染miR-526b-3p阴性对照质粒、miR-526b-3p抑制剂后用30 mmol/L顺阿曲库铵处理。四甲基偶氮唑盐比色法(MTT)检测细胞增殖抑制率;Transwell检测细胞迁移和侵袭;实时荧光定量PCR(RT-qPCR)检测miR-526b-3p和FMNL3 mRNA表达水平;荧光素酶报告实验检测miR-526b-3p和FMNL3的靶向关系。结果:胃癌组织中miR-526b-3p表达水平低于正常组织,FMNL3蛋白及mRNA表达水平高于正常组织(P<0.05)。与对照组相比,顺阿曲库铵低、中、高浓度组胃癌AGS细胞中细胞增殖抑制率升高,迁移和侵袭细胞数降低,miR-526b-3p表达水平升高,FMNL3 mRNA和蛋白表达水平降低,且呈浓度依赖性(P<0.05)。miR-526b-3p过表达抑制胃癌AGS细胞的增殖、迁移和侵袭。miR-526b-3p靶向调控FMNL3的表达,抑制miR-526b-3p表达逆转了顺阿曲库铵对胃癌AGS细胞的抑制作用。结论:顺阿曲库铵可抑制胃癌AGS细胞增殖、迁移和侵袭,其机制可能与miR-526b-3p和FMNL3有关。

Objective:To investigate the effect and mechanism of cisatracurium on the proliferation,migration and invasion of gastric cancer AGS cells.Methods:A total of 30 cases of gastric cancer tissues and corresponding normal gastric mucosal tissues from January 2019 to January 2022 were selected.The experiment was divided into the control group(normal cultured cells),cisatracurium low(10 mmol/L),middle(20 mmol/L),and high(30 mmol/L)concentration group,miR-NC group,miR-526b-3p group,anti-miR-NC group,anti-miR-526b-3p group,Cis+anti-miR-NC group,Cis+anti-miR-526b-3p group.The cells in Cis+anti-miR-NC group,Cis+anti-miR-526b-3p group were transfected with miR-526b-3p negative control plasmid,miR-526b-3p inhibitor and then treated with 30 mmol/L cisatracurium.Cell proliferation inhibition rate was detected by MTT assay.Transwell assay was used to detect cell migration and invasion.Real-time quantitative PCR(RT-qPCR)was used to detect miR-526b-3p and FMNL3 mRNA expression.Luciferase reporter assay was used to detect the targeting relationship of miR-526b-3p and FMNL3.Results:The expression level of miR-526b-3p in gastric cancer tissues was lower than that in normal tissues,and the expression level of FMNL3 protein and mRNA was higher than that in normal tissues(P<0.05).Compared with control group,the cell proliferation inhibition rate in cell treated with cisatracurium low,middle and high concentrations group was increased,the number of migration and invasion cells was decreased,the expression of miR-526b-3p was increased,and the expression of FMNL3 mRNA and protein was significantly decreased ( P <0.05).Overexpression of miR-526b-3p inhibits the proliferation,migration and invasion of AGS cells.miR-526b-3p targeted the regulation of FMNL3 expression,and inhibition of miR-526b-3p expression reversed the inhibitory effect of cisatracurium on AGS cells. Conclusion: Cisatracurium can inhibit the proliferation,migration and invasion of gastric cancer AGS cells,and its mechanism may be related to miR-526b-3p and FMNL3.