Cytochrome P450 ω-hydroxylase promotes angiogenesis and metastasis by upregulation of VEGF and MMP-9 in non-small cell lung cancer

Abstract

Purpose Cytochrome P450 (CYP) ω-hydroxylase, mainly consisting of CYP4A and CYP4F, converts arachidonic acid to 20-hydroxyeicosatetraenoic acid (20-HETE) that induces angiogenic responses in vivo and in vitro. The present study examined the role of CYP ω-hydroxylase in angiogenesis and metastasis of human non-small cell lung cancer (NSCLC). Methods The effect of WIT003, a stable 20-HETE analog, on invasion was evaluated using a modified Boyden chamber in three NSCLC cell lines. A549 cells were transfected with CYP4A11 expression vector or exposed to CYP ω-hydroxylase inhibitor (HET0016) or 20-HETE antagonist (WIT002), and then ω-hydroxylation activity toward arachidonic acid and the levels of matrix metalloproteinases (MMPs) and VEGF were detected. The in vivo effects of CYP ω-hydroxylase were tested in established tumor xenografts and an experimental metastasis model in athymic mice. Results Addition of WIT003 or overexpression of CYP4A11 with an associated increase in 20-HETE production significantly induced invasion and expression of VEGF and MMP-9. Treatment of A549 cells with HET0016 or WIT002 inhibited invasion with reduction in VEGF and MMP-9. The PI3 K or ERK inhibitors also attenuated expression of VEGF and MMP-9. Compared with control, CYP4A11 transfection significantly increased tumor weight, microvessel density (MVD), and lung metastasis by 2.5-fold, 2-fold, and 3-fold, respectively. In contrast, WIT002 or HET0016 decreased tumor volume, MVD, and spontaneous pulmonary metastasis occurrences. Conclusion CYP ω-hydroxylase promotes tumor angiogenesis and metastasis by upregulation of VEGF and MMP-9 via PI3 K and ERK1/2 signaling in human NSCLC cells.

Translational Implications of the Alcohol-Metabolizing Enzymes, Including Cytochrome P450-2E1, in Alcoholic and Nonalcoholic Liver Disease

“…In contrast, the potent vasoconstrictive 20-HETE, which has strong mitogenic and angiogenic properties, is increased in tumors of liver, brain, kidney, and ovary with increased expression of CYP4A/4F genes compared to those in normal tissues (Alexanian, Miller, Roman, & Sorokin, 2012). Similarly, increased expression of CYP4A11, CYP4F2, and CYP4F3 isoforms were significantly expressed in pancreatic ductal adenocarcinoma (Gandhi et al, 2013), suggesting that 20-HETE, which increases expression of HIF and its downstream target vascular endothelial growth factor (VEGF), promotes blood vessel sprouting and metastasis by activation of metalloproteinases (MMPs) (Yu et al, 2011). Thus, selective inhibitors of 20-HETE synthesis by CYP4 omega hydroxylase have been demonstrated to reduce proliferation, angiogenesis, and invasion in lung, renal, and brain cancers (Edson & Rettie, 2013).…”

Impaired myogenic response and autoregulation of cerebral blood flow is rescued in CYP4A1 transgenic Dahl salt-sensitive rat

“…20-HETE has also been reported to play a critical role in angiogenesis (65), restenosis (40), acute renal injury (10,46,49), chronic kidney disease (10), cancer (1), hypertension (10,48,63), and ischemic and hemorrhagic stroke (8,39,47). The availability of the Dahl SS rat model deficient in the formation of 20-HETE and along with our new rescued CYP4A1 transgenic SS rat model also provides investigators with a unique opportunity to study the mechanisms by which 20-HETE contributes to the pathophysiology of these diseases.…”

Vascular actions of 20-HETE

“…CYP ω-hydroxylases facilitate growth and metastasis of human non-small cell lung cancer through PI3K signaling and angiogenic responses [116]. In human non-small cell lung cancer cells, a stable 20-HETE analog (WIT003) or overexpression of CYP4A11 induced invasion of the cells in a modified Boyden chamber assay associated with expression of VEGF and MMP-9 [116]. The induction of VEGF and MMP-9 were blocked by inhibitors of PI3K or ERK signaling.…”