Combined treatments of heat, radiation, or cytokines with flavone acetic acid on the growth of cultured endothelial cells

Abstract

The antitumour effects of flavone acetic acid (FAA) against a broad spectrum of established experimental tumours has been demonstrated. Damage to the vasculature, which rapidly disrupts blood flow and induces haemorrhagic necrosis, is believed to be a major mechanism contributing to the observed antitumour effects. Despite these established observations, FAA has shown little effect against human tumours. However, other applications of FAA, for examples, for an extended period of treatments or in combination with other antitumour modalities, have not been sufficiently explored. In order to test the direct effects of FAA on vasculature, endothelial cells isolated from human umbilical vein (HUVEC) and bovine pulmonary artery (CPAEC) were used in this study. FAA at the concentrations of 50 to 200 μg/ml causes reduction in cell number (from 20 to > 30% of the cells) of HUVEC as measured by MTT assay after 1, 3, and 5 h of treatment at 37°C. FAA did not produce significant effects on similarly treated human squamous cell carinoma, cell line UM-SCC-2. After 1 h treatment of FAA at 300 μg/ml, a large number of HUVECs failed to react with an actin stain, NBD-phallacidin. The growth of HUVECs and CPAEC in the presence of FAA for 1–3 days was progressively reduced. The number of HUVEC treated for 3 days at the concentrations of 100, 200, and 300 μg/d were reduced by 75–86% in comparison with the control culture. The experiments with CPAEC showed similar results. The inhibition of the growth of endothelial cells by FAA was enhanced when it combines with tumour necrosis factor-a but not with interleukin-I, interferon-γ, heat, or radiation. We observed that FAA can initiate both immediate effects and growth inhibition on cultured endothelial cells. These results support the notion that FAA rapidly induces vasculature damage. Furthermore, cytokines such as tumour necrosis factor-α can enhance the toxicity of FAA on endothelial cells.

Key Words:

• Heat
• radiation
• tumour necrosis factor-a
• flavone acetic acid
• endothelial cell