摘要
Objective:To explore the role of circROBO1 in promoting the invasion of retinal Y79 cells by targeting KLF5 and its possible regulatory mechanism.Methods:RNase R enzyme digestion and qRT-PCR experiments were used to detect the structural stability of circular circROBO1 in retinal Y79 cells;cytoplasmic and nuclear RNAs of retinal Y79 cells were extracted for localization analysis of circROBO1;The expression of circROBO1 in retinal Y79 cells were silenced by siRNA.The effect of circROBO1 on the migration and invasion ability of HT-29 cells was detected by scratch assay,Transwell cell invasion and migration assay.The target binding sites of circROBO1 and its downstream miRNA and that of miRNA and its downstream target gene KLF5 were predicted by CircInteractome and TargetScan online software respectively,and the target regulation relationship between them was verified by double luciferase reporter gene experiment.Western blot was used to detect the effect of siRNA silencing the expression of circROBO1 in Y79 cells on the expression of KLF5.Results:Compared with the control group without RNase R enzyme treatment,relative circROBO1 levels did not change significantly after treatment,while relative linear ROBO1 levels decreased significantly after treatment(t=16.18,P<0.05);the content of circROBO1 in the cytoplasm was significantly higher than that in the nucleus(P<0.05);compared with si-control group,the migration rate and the invasion and migration abilities of Transwell cells were all lower in the si-circROBO1 group(t=22.54,P<0.05);circROBO1 can adsorb miR-885-5p,and there is a target binding site between miR-885-5p and KLF5(t=11.39,P<0.05);compared with the si-control group,the KLF5 protein expression in the si-circROBO1 group was significantly decreased(t=17.26,P<0.05).Conclusions:circROBO1 promotes retinalY79 cell tumor invasion by targeting KLF5.
基金
Key Project of Hunan Provincial Department of Education(No.21A0285)。
