CircRNA EPB41L2 inhibits proliferation,migration and invasion of oral squamous carcinoma cells by regulating the microRNA-1270/GNAI3 axis
YANG Feng
MIAO Ke-hong
LAN Yu-yan
LIN Fang-liang
SUN Li-bo
Abstract：Objective 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 mRNA 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 significantly decreased,while the apoptosis rate was significantly increased(P＜0.001).Compared with the oe EPB41 L2+mimics NC group,the proliferation ability,scratch healing rate and invasion number of CAL-27 cells in oe EPB41L2+miR mimics group were significantly increased,while the apoptosis rate was significantly decreased(P＜0.001).Compared with HOK group,the expression of miR-1270 in CAL-27 group was significantly increased(P＜0.001).Compared with the oe NC group,the expression of miR-1270 in the oe EPB41L2 group was significantly decreased(P＜0.001).Compared with HOK group,the expression of GNAI3 mRNA and protein in CAL-27 group was significantly decreased(P＜0.001).Compared with oe NC group,the expression of GNAI3 mRNA and protein in oe EPB41L2 group was significantly increased(P＜0.001).Compared with oe EPB41L2+mimics NC group,the expression of GNAI3 mRNA and protein in oe EPB41L2+miR mimics group was significantly decreased(P＜0.05).RNA pull-down assay,RIP assay and dual luciferase reporting assay jointly confirmed the targeting binding of miR-1270 to circRNA EPB41L2.Dual luciferase assay confirmed the targeting of miR-1270 to GNAI3.Conclusion circRNA EPB41L2 inhibited the proliferation,migration and invasion of OSCC cells by regulating the miR-1270/GNAI3 axis.
Keywords：oral squamous cell carcinomacircular RNA EPB41L2microRNA-1270GNAI3proliferationmigration
Publication Date：2025-01-20
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：8( 4-11 )