Inhibition of hemangioma development by regulating the VEGF/VEGFR autocrine loop via the miR-494/PTEN pathway.

Abstract

Background: Infantile hemangioma (IH) is the most common type of benign vascular tumor found in infants and young children. Hemangioma-derived endothelial cells within the lesion from birth to three months of age are the primary characteristic of IH. (hemangioma-derived endothelial cells, HemECs) proliferated rapidly and formed hemangioma masses, most of which gradually regressed spontaneously within the next 1 to 5 years of age and continued to improve until the age of 6 to 12 years. But 10-15% of IH cases can still result in ulcerative, obstructive, deformity, and even potentially fatal consequences and sequelae. These conditions seriously affect children's physical and mental health as well as their growth and development, necessitating prompt and efficient medical attention of IH is known.

Objective: The purpose of this work. HemECs are crucial to the development of IH as a result. Not all of the pathophwork is to examine the impact of OMT on HemECs, with a specific emphasis on its role in cell migration, proliferation, cell cycle regulation, and apoptosis. Additionally, we will research the influence of OMT on the VEGFA/VEGFR-2 signaling pathway in HemECs and assess the impact of OMT on the miR-494/PTEN axis.

Methods: The Cell Counting Kit-8 (CCK-8) assay was employed to evaluate the influence of Oxymatrine (OMT) on the proliferation of Hemangioma Endothelial Cells (HemECs). The Transwell Assay was employed to detect cell invasion and migration. The cell cycle and apoptosis were analyzed through flow cytometry. The impact of OMT on the expression of apoptosis markers (cleaved caspase-3) and proteins associated with the cell cycle (Cyclin D1, Bcl-2, Bax) was examined using Western Blot and Reverse Transcription Polymerase Chain Reaction (RT-PCR).

Results: OMT treatment significantly inhibited the proliferation of HemECs, especially when combined with the miR-494 inhibitor. Additionally, OMT administration raised the proportion of cells entering the G2 phase, accelerated apoptosis, and decreased HemECs' capacity for migration and invasion. The results of Western Blot and RT-PCR demonstrated that OMT decreased the expression levels of Bax and Cleaved Caspase-3 while increasing the expression of Bcl-2 and Cyclin D1. OMT and miR-494 inhibitors have distinct impacts on the phosphorylated versions of VEGFR-2, PTEN, and ERK signaling pathways. OMT may control cell survival and proliferation through the miR-494/PTEN pathway, as evidenced by the fact that PTEN expression was dramatically upregulated in the miR-1297 inhibitor with OMT treatment group, while p-ERK expression was markedly reduced in that group.

Conclusions: OMT effectively inhibits the growth, migration, and apoptosis of hemangioma endothelial cells, likely by regulating key proteins involved in the cell cycle and apoptosis. The combination with miR-494 inhibitors enhances its therapeutic effect, suggesting a potential new approach for hemangioma treatment. These findings support the development of OMT-based strategies for hemangiomas and its potential use in cancer therapy.