HET0016, a Selective Inhibitor of 20-HETE Synthesis, Decreases Pro-Angiogenic Factors and Inhibits Growth of Triple Negative Breast Cancer in Mice

Abstract

A selective inhibitor of 20-HETE synthesis, HET0016, has been reported to inhibit angiogenesis. 20-HETE has been known as a second mitogenic messenger of angiogenesis inducing growth factors. HET0016 effects were analyzed on MDA-MB-231 derived breast cancer in mouse and in vitro cell line. MDA-MB-231 tumor cells were implanted in animals’ right flank and randomly assigned to early (1 and 2), starting treatments on day 0, or delayed groups (3 and 4) on day 8 after implantation of tumor. Animals received HET0016 (10 mg/kg) treatment via intraperitoneal injection for 5 days/week for either 3 or 4 weeks. Control group received vehicle treatment. Tumor sizes were measured on days 7, 14, 21, and 28 and the animals were euthanized on day 22 and 29. Proteins were extracted from the whole tumor and from cells treated with 10 µM HET0016 for 4 and 24 hrs. Protein array kits of 20 different cytokines/factors were used. ELISA was performed to observe the HIF-1α and MMP-2 protein expression. Other markers were confirmed by IHC. HET0016 significantly inhibited tumor growth in all treatment groups at all-time points compared to control (p<0.05). Tumor growth was completely inhibited on three of ten animals on early treatment group. Treatment groups showed significantly lower expression of pro-angiogenic factors compared to control at 21 days; however, there was no significant difference in HIF-1α expression after treatments. Similar results were found in vitro at 24 hrs of HET0016 treatment. After 28 days, significant increase of angiogenin, angiopoietin-1/2, EGF-R and IGF-1 pro-angiogenic factors were found (p<0.05) compared to control, as well as an higher intensity of all factors were found when compared to that of 21 day’s data, suggesting a treatment resistance. HET0016 inhibited tumor growth by reducing expression of different set of pro-angiogenic factors; however, a resistance to treatment seemed to happen after 21 days.

Molecular mechanisms in differentiated thyroid cancer

“…This was associated with stimulation of ROS production, the secretion of HIF-1alpha and activation of the PI3K/AKT pathway (185, 198, 205). In addition, administration of HET0016 has been reported to decrease the expression of a variety of angiogenic growth factors and inhibit growth of triple negative breast cancer tumor in mice (206). In human small cell lung cancer cell line, treatment with the 20-HETE agonist, 20-hydroxyeicosa-5(Z),14(Z)-dienoic acid (WIT003) or upregulation of the expression of CYP4A11, induced expression of VEGF and MMP-9 via PI3K or ERK pathways that significantly increased microvascular density.…”

“…The findings that the expression of CYP4A/4F enzymes are elevated in human thyroid, ovarian, breast, and colon cancers (211), pancreatic adenocarcinoma (212) further support that 20-HETE may play a role in vascularization and growth of cancerous tumors. More recent studies indicate that inhibition of the synthesis of 20-HETE are effective in reducing the growth of glioblastoma, lung, prostate, renal, breast and colon cancer in experimental animals (206, 213–217). …”

Chemical synthesis and biological evaluation of ω-hydroxy polyunsaturated fatty acids

“…20-HETE has been shown to have detrimental effects in several diseases such as hypertension, 8 cancer, 9 and cardiovascular and kidney diseases, 10 despite its low in vivo concentrations due to reincorporation into membrane phospholipid pools, and plasma protein binding similar to other fatty acids. 1 In addition, significantly increased 20-HETE resulting from chronic administration of rofecoxib to mice suggests that it may contribute to the cardiovascular risks associated with coxibs and nonselective nonsteroidal anti-inflammatory drugs (NSAIDs).…”

Shexiang Baoxin pills promotes angiogenesis in myocardial infarction rats via up-regulation of 20-HETE-mediated endothelial progenitor cells mobilization

“…Rats in MI+ ShexiangBaoxin pills group received a gavage of ShexiangBaoxin pills (25 mg/kg/d) [25] in distilled water for 8 weeks. HET0016 was given through intraperitoneal injection (10 mg/kg/day) [26]. ShexiangBaoxin pills was bought from Shanghai Hutchison Pharmaceuticals.…”