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Abstract
Melanoma is a serious malignant form of skin cancer. Euphorbiaceae compound B (ECB, 2,4-dihydroxy-6-methoxy-3-methylacetophenone) is an acetophenone compound that is isolated from Euphorbia ebracteolata Hayata (EEH), an herbaceous perennial, and has antitumor activity. Here, we transplanted human melanoma cells into zebrafish embryos to establish a zebrafish/melanoma model. We showed that this model can be used to evaluate the therapeutic effect of EEH and ECB and discussed its potential mechanism of action. The results showed that ECB was an active ingredient of EEH in inhibiting melanoma-induced hyperplasia of blood vessels in zebrafish embryos, similar to the angiogenic inhibitor vatalanib. ECB inhibited the number and length of subintestinal veins (p < 0.05), as well as the distribution of melanoma in zebrafish embryos (p < 0.05). More importantly, unlike vatalanib, ECB only inhibited melanoma-induced abnormal and excessive growth of blood vessels in xenografts. In addition, ECB inhibited the mRNA expression of vegfr2 and vegfr3 in zebrafish. Both vegfr2 and vegfr3 are essential genes that regulate blood vessel formation and upregulate the expression of p53 and casp3a genes in zebrafish. Together, the above-mentioned results indicate that ECB has a potential antimelanoma effect in vivo, which may be mediated by inhibiting vascular endothelial growth factor receptors.
Highlights
Malignant melanoma is the most serious type of skin cancer, which is caused by hyperplasia of melanocytes in the skin
At 24 hpi, 50 μg/mL of ebracteolata Hayata (EEH) was added to the embryo medium of the zebrafish/melanoma xenograft, containing the injected cells labeled with red fluorescent dye (CM-DiI, Invitrogen)
Tumor-induced angiogenesis was quantified by evaluating the number and length of ectopic vessels in the subintestinal vein (SIV) of zebrafish/tumor xenografts at 24 and 48 h after injection (24 and 48 hpi) (Figure 1). e length of the SIV ectopic blood vessels in control Human umbilical vein endothelial cells (HUVECs) and A2058 cells was measured
Summary
Malignant melanoma is the most serious type of skin cancer, which is caused by hyperplasia of melanocytes in the skin. Dihydroxy-6-methylbenzoidone) is a compound of benzene ketones in the Euphorbia ebracteolata Hayata (EEH) and Euphorbia fischeriana Steud [2, 3]. Studies that focused on the anticancer effects of diterpenoids in Euphorbiaceae have been reported; studies on ECB are limited. Blocking the pathway underlying tumor angiogenesis may be a unique way to block the growth of blood vessels that. Evidence-Based Complementary and Alternative Medicine support tumor growth [4]. E zebrafish model can be used as a viable model for whole-animal screening using small molecules that affect tumor angiogenesis [5]. The subintestinal vein (SIV) of zebrafish/tumor xenografts can be visualized noninvasively in real time using vascularspecific transgenic zebrafish or can be stained using blood vessel dye and visualized under a microscope [6]. Koenig et al showed that vegfaa was expressed at the site of intestinal vascularization and might provide a guiding signal and that vegfc overexpression could lead to SIV overgrowth
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