Upregulation of AKT1 and downregulation of AKT3 caused by dysregulation of microRNAs contributes to pathogenesis of hemangioma by promoting proliferation of endothelial cells
Author(s) -
Lu Shuo,
Chen Lingling,
Tang Li
Publication year - 2019
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.28741
Subject(s) - downregulation and upregulation , microrna , transfection , akt1 , pathogenesis , cancer research , gene silencing , western blot , hemangioma , cell growth , microbiology and biotechnology , biology , signal transduction , immunology , medicine , cell culture , pathology , gene , pi3k/akt/mtor pathway , genetics
Abstract This study aimed to verify the differentially expressed miRNAs (microRNAs) in hemangioma, and explore their roles in the pathogenesis of hemangioma in vivo and ex vivo. Real‐time polymerase chain reaction (PCR) and western blot were used to measure reported differentially expressed miRNAs and their potential targets. In‐silicon analysis and luciferase assay were conducted to find the target of miR‐15a and miR‐205. The 3‐(4,5‐dimethylthiazol‐2‐yl)‐2,5‐diphenyl tetrazolium bromide (MTT) assay and flowcytometry were performed to examine the effect of dysregulation of miR‐15a and miR‐205 on the proliferation and apoptosis of endothelial cells. Among all candidate miRNAs, only miR‐205 level was significantly downregulated whereas miR‐15a was evidently upregulated in the hemangioma group. Accordingly, AKT3 was validated to be the direct target of miR‐15a and miR‐205. Using real‐time PCR, the level of AKT1 was much higher in hemangioma group, whereas level of AKT3 was much lower in the hemangioma group, and in general expression level of ATK was upregulated in the hemangioma group. Furthermore, the ATK1 level of cells transfected with miR‐205 mimics and ATK1 siRNA was substantially downregulated, and anti‐miR‐205 mimic significantly improved the level of AKT1, and meanwhile the level of ATK3 and PTEN were remarkably suppressed after transfection with miR‐15a mimics and ATK3 siRNA, whereas notably overexpressed after introduction of anti‐miR‐15a. And miR‐15a, AKT3 siRNA and anti‐miR‐205 evidently induced viability, and miR‐205, AKT1 siRNA, and anti‐miR‐15a obviously promoted apoptosis of cells. Conclusion miR‐15a and miR‐205 had different expression in hemangioma, may be novel therapeutic targets in the treatment of hemangioma by targeting AKT3 and AKT1.