miR-370-3p对口腔鳞癌细胞SCC-9增殖、迁移和侵袭的影响  

Effect of miR-370-3p on proliferation,migration and invasion of oral squamous cell carcinoma SCC-9 cells

作　　者：万紫微 粟小平 韦珍夏 黄旋平[1] WAN Ziwei;SU Xiaoping;WEI Zhenxia;HUANG Xuanping(Department of Oraland Maxillofacial Surgery,College of Stomatology,Hospital of Stomatology,Guangxi Medical University,Guangxi Key Laboratory of Oral and Maxillofacial Rehabilitation and Reconstruction,Guangxi Clinical Research Center for Craniofacial Deformity,Nanning 530021,China)

机构地区：[1]广西医科大学口腔医学院·附属口腔医院颌面外科广西口腔颌面修复与重建研究自治区级重点实验室广西颅颌面畸形临床医学研究中心,南宁530021

出　　处：《广西医科大学学报》2024年第1期32-39,共8页Journal of Guangxi Medical University

基　　金：国家自然科学基金资助项目(No.82360187);广西科技基地和人才专项(No.2021AC18031);广西医疗卫生适宜技术开发与推广应用项目(No.S2021085);南宁市青秀区科技计划项目(No.2021004)。

摘　　要：目的:探究miR-370-3p对口腔鳞癌细胞(SCC-9)增殖、迁移和侵袭的影响,并分析其可能的作用机制。方法:比较人正常口腔鳞状上皮细胞(NOK)与SCC-9细胞miR-370-3p和NPC1表达。将SCC-9细胞分为miR-370-3p mimic组、miR-370-3p阴性对照(NC)组、miR-370-3p inhibitor组、miR-370-3p inhibitor-NC组。采用克隆形成实验检测细胞集落形成能力,CCK-8法、划痕实验、Transwell实验分别检测细胞增殖、迁移和侵袭能力,实时荧光定量PCR(RT-qPCR)法检测细胞miR-370-3p表达,双荧光素酶实验检测miR-370-3p与NPC1的靶向关系,western blotting法检测NPC1蛋白表达。结果:与NOK细胞比较,SCC-9细胞miR-370-3p表达上调,NPC1表达下调(均P<0.05)。与mimic NC组比较,miR-370-3p mimic组miR-370-3p表达水平升高,NPC1蛋白表达水平降低,细胞活力增强,划痕迁移率降低,穿膜细胞数增加(均P<0.05)。miR-370-3p inhibitor组上述指标变化趋势与miR-370-3p mimic组相反(均P<0.05)。荧光素酶实验结果显示,miR-370-3p能够靶向结合NPC1的3’-UTR,并抑制NPC1蛋白表达(P<0.05)。结论:miR-370-3p可促进口腔鳞癌细胞SCC-9增殖、迁移和侵袭,其机制可能与调控其下游靶基因NPC1的表达有关。Objective:To explore the effect of miR-370-3p on the proliferation,migration,and invasion of oral squamous cell carcinoma SCC-9 cells,and analyze its possible mechanism of action.Methods:The expression of miR-370-3p and NPC1 in normal human oral keratinocytes(NOK)and SCC-9 cells was compared.SCC-9 cells were divided into miR-370-3p mimic group,miR-370-3p negative control(NC)group,miR-370-3p inhibitor group and miR-370-3p inhibitor-NC group.Clonal formation assay was used to detect cell colony forming ability.Cell counting kit-8(CCK-8)assay,scratch assay and Transwell assay were used to detect cell proliferation,migration and invasion ability,respectively.Real-time fluorescence quantitative PCR(RT-qPCR)assay was used to detect miR-370-3p expression.Dual luciferase assay was used to detect the targeting of miR-370-3p to NPC1,and western blotting was used to detect the expression of NPC1 protein.Results:Compared with NOK cells,the expression of miR-370-3p was up-regulated and the expression of NPC1 was down-regulated in SCC-9 cells(all P<0.05).Compared with the mimic NC group,the expression level of miR-370-3p was increased and the expression level of NPC1 protein was decreased in the miR-370-3p mimic group.Cell viability was enhanced,scratch mobility was decreased,and the number of transmembrane cells was increased(all P<0.05).The change trend of the above indexes of the miR-370-3p inhibitor group was opposite to that of the miR-370-3p mimic group(all P<0.05).Luciferase assay results showed that miR-370-3p could target the 3’-UTR binding to NPC1 and inhibit the expression of NPC1 protein(P<0.05).Conclusion:The miR-370-3p can promote the proliferation,migration and invasion of SCC-9 cells,and its mechanism may be related to the regulation of the expression of its downstream target gene NPC1.

关 键 词：miR-370-3p 口腔鳞状细胞癌 增殖 迁移 侵袭 

分 类 号：R739.8[医药卫生—肿瘤]