环状RNA EPB41L2通过调控微小RNA-1270/GNAI3轴抑制口腔鳞状细胞癌细胞的增殖和迁移及侵袭  

Circ RNA EPB41L2 inhibits proliferation,migration and invasion of oral squamous carcinoma cells by regulating the microRNA-1270/GNAI3 axis

作　　者：杨凤[1] 缪克红[1] 兰玉燕 林方梁 孙黎波 YANG Feng;MIAO Ke-hong;LAN Yu-yan;LIN Fang-liang;SUN Li-bo(Department of Stomatology,First People's Hospital of Zigong City,Zigong 643000,China)

机构地区：[1]自贡市第一人民医院口腔科,643000 [2]西南医科大学口腔修复科,泸州646000 [3]泸州市人民医院口腔科,泸州市646000

出　　处：《中华老年口腔医学杂志》2025年第1期4-11,共8页Chinese Journal of Geriatric Dentistry

摘　　要：目的环状RNA(circular RNA,circRNA)与口腔鳞状细胞癌(oral squamous cell carcinoma,OSCC)的进展密切相关,现探讨circRNA EPB41L2调控OSCC细胞增殖、迁移和侵袭的作用机制。方法将细胞分为7组:HOK组(正常培养的口腔上皮细胞)、SCC-9组(正常培养的SCC-9细胞)、CAL-27组(正常培养的CAL-27细胞)、oe NC组(转染空载pcDNA3.4至CAL-27细胞)、oe EPB41L2组(转染pcDNA3.4-EPB41L2至CAL-27细胞)、oe EPB41L2+mimics NC组(转染pcDNA3.4-EPB41L2及mimics NC至CAL-27细胞)、oe EPB41L2+miR mimics组(转染pcDNA3.4-EPB41L2及miR-1270 mimics至CAL-27细胞)。通过实时荧光定量聚合酶链式反应检测circRNA EPB41L2、miR-1270及GNAI3 mRNA在各组细胞中的表达,通过CCK-8检测各组细胞活力,通过流式细胞术检测各组细胞凋亡率,通过集落形成实验检测各组细胞增殖能力,通过细胞划痕实验检测各组细胞划痕愈合率,通过Transwell实验检测各组细胞侵袭数量,通过Western blot检测GNAI3蛋白在各组细胞中的表达,通过RNA pull-down实验、RNA免疫共沉淀(RNA immunoprecipitation,RIP)实验、双荧光素酶报告实验验证miR-1270与circRNA EPB41L2的靶向结合关系,通过双荧光素酶报告实验验证miR-1270与GNAI3的靶向结合关系。结果与HOK组比较,SCC-9组、CAL-27组中circRNA EPB41L2表达显著下降(P<0.01),且在CAL-27组中表达水平最低,因此选取CAL-27细胞作为后续研究对象。与oe NC组比较,oe-circRNA EPB41L2组CAL-27细胞活力、增殖能力、划痕愈合率、侵袭数量显著下降,细胞凋亡率显著上升(P<0.001)。与oe EPB41L2+mimics NC组比较,oe EPB41L2+miR mimics组CAL-27细胞增殖能力、划痕愈合率、侵袭数量显著上升,细胞凋亡率显著下降(P<0.001)。与HOK组比较,CAL-27组细胞miR-1270表达显著升高(P<0.001);与oe NC组比较,oe EPB41L2组细胞miR-1270表达显著下降(P<0.001)。与HOK组比较,CAL-27组细胞GNAI3 mRNA和蛋白表达显著下降(P<0.001);与oe NC组比较,oe EPB41L2组细胞GNAIObjective circular RNA(circRNA)is closely related tothe progression of oral squamous cell carcinoma(OSCC),the aim of this study was to investigate the mechanism of action of circRNA EPB41L2 in regulating the proliferation,migration and invasion of OSCC cells.Methods The cells were divided into seven groups:HOK group(normal cultured oral epithelial cells),SCC-9 group(normal cultured SCC-9 cells),CAL-27 group(normal cultured CAL-27 cells),oe NC group(CAL-27 cells transfected with empty pcDNA3.4),oe EPB41L2 group(CAL-27 cells transfected with pcDNA3.4-EPB41L2),oe EPB41L2+mimics NC group(transfected with pcDNA3.4-EPB41L2 and mimics NC to CAL-27 cells),oe EPB41L2+miR mimics group(transfected with pcDNA3.4-EPB41L2 and miR-1270 mimics to CAL-27 cells).Real-time fluorescence quantitative polymerase chain reaction was used to detect the expression of circRNA EPB41L2,miR-1270 and GNAI3 mR NA in the cells of each group,CCK-8 was used to detect cell viability,flow cytometry was used to detect cell apoptosis,and colony formation assay was used to detect cell proliferation.The scratch healing rate of cells in each group was detected by cell scratch experiment,the number of cell invasion in each group was detected by Transwell experiment,and the expression of GNAI3 protein in cells in each group was detected by Western blot.The targeted binding relationship between miR-1270 and circRNA EPB41L2 was verified through RNA pull-down assay,RNA immunoprecipitation(RIP)assay and dual luciferase reporting assay.The targeting binding relationship between miR-1270 and GNAI3 was verified by double luciferase reporting assay.Results Compared with HOK group,the expression of circRNA EPB41L2 in SCC-9 group and CAL-27 group was significantly decreased(P<0.01),and the expression level was lowest in CAL-27 group,.Ttherefore,CAL-27 cells were selected as the object of follow-up study.Compared with the OE-NC group,the viability,proliferation ability,scratch healing rate and invasion number of CAL-27 cells in the oe-circRNA EPB41L2 group were signi

关 键 词：口腔鳞状细胞癌 环状RNA EPB41L2 微小RNA-1270 GNAI3 增殖 迁移 

分 类 号：R739.8[医药卫生—肿瘤]