微小RNA-590对口腔鳞癌细胞增殖、迁移、侵袭的影响及其机制

Effect and mechanism of miR-590 on the proliferation,migration and invasion of oral squa⁃mous cell carcinoma cells

目的探讨微小RNA(miR)-590对口腔鳞癌细胞增殖、迁移、侵袭的影响和分子机制。方法实时荧光定量逆转录聚合酶链反应(qRT-PCR)检测2014年1月至2017年1月菏泽市牡丹人民医院52例口腔鳞癌组织中miR-590的表达。将口腔鳞癌细胞Tca-8113分为miRNA抑制物阴性对照(anti-miR-NC)组、miR-590抑制物(anti-miR-590)组、空载体(pcDNA3.1)组、大肿瘤抑制基因1(LATS1)过表达载体(pcDNA3.1-LATS1)组、anti-miR-590+小干扰RNA阴性对照(si-NC)组、anti-miR-590+LATS1小干扰RNA组(si-LATS1)。四甲基偶氮唑盐微量酶反应比色法(MTT法)检测细胞活力,Transwell法检测细胞的迁移和侵袭数。双荧光素酶报告基因实验和蛋白质印迹法检测验证miR-590和LATS1的靶向调控关系。结果与癌旁组织比较,口腔鳞癌组织中miR-590的表达水平显著升高[(0.73±0.07)比(0.30±0.03),P<0.05]。与anti-miR-NC组比较,anti-miR-590组细胞活力[(0.40±0.04)比(0.78±0.08)]、迁移[(46±5)个比(136±13)个]和侵袭数[(42±4)个比(128±13)个]显著降低(P<0.05);与pcD⁃NA3.1组比较,pcDNA3.1-LATS1组细胞活力[(0.49±0.05)比(0.78±0.08)]、迁移数[(56±6)个比(136±13)个]和侵袭数[(50±5)个比(128±13)个]显著降低(P<0.05)。LATS1靶向负调控LATS1表达。与anti-miR-590+si-NC组比较,anti-miR-590+si-LATS1组细胞活力(0.61±0.06)比(0.40±0.04)、迁移数[(94±9)个比(45±4)个]和侵袭数[(87±9)个比(40±4)个]显著升高(P<0.05)。结论抑制miR-590通过靶向调控LATS1表达从而抑制口腔鳞癌细胞的增殖、迁移和侵袭。

Objective To explore the effects and molecular mechanisms of microRNA(miR)-590 on the proliferation,migration and in⁃vasion of oral squamous cell carcinoma cells.Methods Real-time fluorescent quantitative reverse transcription polymerase chain reac⁃tion(qRT-PCR)was used to detect the expression of miR-590 in 52 cases of oral squamous cell carcinoma tissues from Mudan People's Hospital in Heze City from January 2014 to January 2017.Oral squamous cell carcinoma cells Tca-8113 were divided into miRNA in⁃hibitor negative control(anti-miR-NC)group,miR-590 inhibitor(anti-miR-590)group,empty vector(pcDNA3.1)group,and large tu⁃mor suppressor gene 1(LATS1)overexpression vector(pcDNA3.1-LATS1)group,anti-miR-590+small interfering RNA negative control(si-NC)group,anti-miR-590+LATS1 small interfering RNA group(si-LATS1)).Methyl thiazolyl tetrazolium(MTT)method detected cell viability,and the Transwell method detected cell migration and invasion.The dual luciferase reporter gene experiment and western blotting test were applied to verify the targeted regulation relationship between miR-590 and LATS1.Results Compared with adjacent tissues,the expression level of miR-590 in oral squamous cell carcinoma tissues was significantly higher[(0.73±0.07)vs.(0.30±0.03),P<0.05].Compared with the anti-miR-NC group,the cell viability(0.40±0.04)vs.(0.78±0.08),migration[(46±5)vs.(136±13)]and inva⁃sion number[(42±4)vs.(128±13)]in anti-miR-590 group were significantly lower(P<0.05);compared with the pcDNA3.1 group,the cell viability[(0.49±0.05)vs.(0.78±0.08)],migration[(56±6)vs.(136±13)]and invasion numbers[(50±5)vs.(128±13)]in pcDNA3.1-LATS1 group were significantly lower(P<0.05);LATS1 targets and negatively regulates LATS1 expression.Compared with the antimiR-590+si-NC group,the cell viability[(0.61±0.06)vs.(0.40±0.04)],migration[(94±9)vs.(45±4)]and invasion numbers[(87±9)vs.(40±4)]in anti-miR-590+si-LATS1 group were significantly higher(P<0.05).Conclusion Inhibition of miR-590 could inhibit the pro⁃liferation,migration and invasion of oral squamous cell carcinoma by targeting the regulation of LATS1 expression.