Propranolol induces hemangioma endothelial cell apoptosis via a p53‑BAX mediated pathway

Abstract

The use of propranolol for the treatment of infantile hemangioma (IH) has been widely investigated in recent years. However, the underlying therapeutic mechanism of propranolol for the treatment of IH remains poorly understood. The aim of the present study was to investigate the expression of proteins regulated by cellular tumor antigen p53 (p53) in associated apoptosis pathways in IH endothelial cells (HemECs) treated with propranolol. Furthermore, the present study aimed to investigate the exact apoptotic pathway underlying the therapeutic effect of propranolol against IH. In the present study, HemECs were subcultured and investigated using an inverted phase contrast microscope, immunocytochemical staining and a scanning electron microscope (SEM). Experimental groups and blank control groups were prepared. All groups were subjected to drug treatment. A high p53 expression model of HemECs was successfully established via transfection, and a low p53 expression model of HemECs was established using pifithrin-α. The apoptosis rate of each group was determined using Annexin V-fluorescein isothiocyanate/propidium iodide double staining and flow cytometry. The expression levels of downstream proteins regulated by p53 [tumour necrosis factor receptor superfamily member 6 (FAS), p53-induced death domain-containing protein (PIDD), death receptor 5 (DR5), BH3-interacting domain death agonist (BID), apoptosis regulator BAX (BAX), p53 unregulated modulator of apoptosis (PUMA), phosphatidylinositol-glycan biosynthesis class S protein (PIGS), and insulin-like growth factor-binding protein 3 (IGF-BP3)] were revealed in the experimental and control groups via western blotting. Microscopic observation revealed the growth of an adherent monolayer of cells, which were closely packed and exhibited contact inhibition. Immunocytochemical staining demonstrated increased expression of clotting factor VIII. SEM analysis revealed presence of Weibel-Palade bodies. The results of the analyses verified that the cultured cells were HemECs. The staining of the samples resulted in a significantly increased rate of apoptosis in experimental groups compared with the blank control group. This result suggested that there is an association between p53 expression and the rate of apoptosis of propranolol-treated HemECs. The results of the western blot analysis demonstrated an upregulation of BAX expression and a downregulation of IGF-BP3 expression in the HemECs treated with propranolol. There were no significant differences in the expression levels of FAS, DR5, PIDD, BID, PUMA and PIGS between experimental and control groups. This result suggests that p53 has an important role in HemEC apoptosis. The results of the present study additionally suggest that the propranolol-induced HemEC apoptosis pathway is a mitochondrial apoptosis pathway and is regulated by p53-BAX signaling.

Topical Timolol Vs. Oral Propranolol for the Treatment of Superficial Infantile Hemangiomas

“…According to recent studies focusing on the potential mechanisms of different anti-hemangioma drugs, propranolol and other beta-blockers mainly exert their effects via targeting hemangioma endothelial cells and hemangioma pericytes (30). Bischoff et al proposed that propranolol could suppress the development of hemangiomas through increasing the contractility of hemangioma pericytes (31), and several other studies reported that propranolol could inhibit the growth of hemangiomas through modulating cellular functions of hemangioma endothelial cells (32–34). Although no significant differences in the therapeutic efficacy of propranolol group and timolol group in our study were observed, we hypothesized that systemic propranolol and local timolol might individually exerted their effects partly via targeting different cells.…”

Integrated WGCNA and PPI Network to Screen Hub Genes Signatures for Infantile Hemangioma

“…Promoting cell cycle arrest and apoptosis are the main mechanisms of action of drugs for the treatment of IH ( Chen et al, 2019 ). In particular, p53 signaling pathway mediated apoptosis ( Yao et al, 2018 ). The mammalian rapamycin complex (mTOR), which converts signals in the extracellular environment, such as glucose and growth factors, into corresponding changes in basic intracellular processes, including proliferation ( Laplante and Sabatini, 2012 ).…”

Huanglian decoction suppresses the growth of hepatocellular carcinoma cells by reducing CCNB1 expression

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