Propranolol regulates the proliferation of hemangioma endothelial cells by targeting microRNA-194-5p with long non-coding RNA nuclear paraspeckle assembly transcript 1
Objective To investigate the role of propranolol on the proliferation of hemangioma-de-rived endothelial cells(HemECs)by targeting microRNA(miR)-194-5p expression with long non-coding RNA nuclear paraspeckle assembly transcript 1(lncRNA NEAT1).Methods Totally,20 cases of hemangioma specimens were surgically removed and collected in the Department of Pediatric Surgery of the Second Hospital of Hebei Medical University from March 2021 to August 2022.The specimens were col-lected and divided into proliferative phase and involuting phase according to Mulliken criteria.The levels of NEAT1 and miR-194-5p in tumors were detected by real-time quantitative reverse transcription polymerase chain reaction(RT-qPCR).HemECs were prepared and the effects of knockdown and overexpression of NEAT1 on the proliferation of HemECs were determined by cell counting kit-8(CCK-8)assay.The inter-action between miR-194-5p and NEAT1 was verified by dual-luciferase assay.After different concentrations of propranolol were administered,the cell activity was detected by CCK-8 assay,cell proliferation was de-tected by 5-Ethynyl-2'-deoxyuridine(EdU)assay,and NEAT1 and miR-194-5p were detected by RT-qPCR.The independent samples t-test was used for comparison between the two groups.The one-way ANOVA and repeated-measures ANOVA were used for comparisons between multiple groups,and Spearman's analysis was used to determine the correlation between the indicators.Results RT-qPCR showed that the expression of NEAT1 in proliferative hemangiomas was higher than that in involuting he-mangiomas(17.43±5.38 vs.8.82±3.70,t=4.176,P＜0.01).Contrarily,the expression of miR-194-5p was decreased(0.28±0.11 vs.0.41±0.11,t=2.692,P＜0.01).CKK-8 assay showed that the cell activity of si-NEAT1 group was lower than that of the si-NC group(0.86±0.05 vs.1.11±0.03,t=7.794,P＜0.01);whereas the cell activity of OE-NEAT1 group was higher than that of the OE-NC group(1.33±0.01 vs.1.11±0.05,t=8.072,P＜0.05).The expression level of miR-194-5p in si-NEAT1 group was higher than that in si-NC group(2.88±0.33 vs.1.14±0.47,t=5.206,P＜0.01);whereas the expression level of miR-194-5p in the OE-NEAT1 group was lower than that in the OE-NC group(0.43±0.06 vs.1.24±0.11,t=10.830,P＜0.01).Dual-luciferase assay showed that luciferase activity was higher in the miR-194-5p and NEAT1-WT co-transfection groups than in the miR-NC and NEAT1-WT co-transfection groups(0.35±0.05 vs.0.65±0.04,t=5.273,P＜0.01).The EdU staining positive rate of cells in the OE-NEAT1 plus propranolol group was significantly lower than that in the OE-NEAT1 and OE-NC groups(35.97±1.09 vs.69.66±2.85,51.89±1.09,F=65.100,P＜0.01).The expression levels of NEAT1 and miR-194-5p in OE-NEAT1 plus propranolol group were signif-icantly different from those in OE-NEAT1 group(1.63±0.30 vs.3.94±0.50,t=6.851,P＜0.01;0.86±0.26 vs.0.37±0.03,t=3.213,P＜0.05).Conclusion Propranolol can regulate the prolifera-tion of HemECs by targeting miR-194-5p via NEAT1,and NEAT1 may be a potential target for molecular therapy of infantile hemangioma.
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