Increased expression of CYP4Z1 promotes tumor angiogenesis and growth in human breast cancer

Abstract

Cytochrome P450 (CYP) 4Z1, a novel CYP4 family member, is over-expressed in human mammary carcinoma and associated with high-grade tumors and poor prognosis. However, the precise role of CYP4Z1 in tumor progression is unknown. Here, we demonstrate that CYP4Z1 overexpression promotes tumor angiogenesis and growth in breast cancer. Stable expression of CYP4Z1 in T47D and BT-474 human breast cancer cells significantly increased mRNA expression and production of vascular endothelial growth factor (VEGF)-A, and decreased mRNA levels and secretion of tissue inhibitor of metalloproteinase-2 (TIMP-2), without affecting cell proliferation and anchorage-independent cell growth in vitro. Notably, the conditioned medium from CYP4Z1-expressing cells enhanced proliferation, migration and tube formation of human umbilical vein endothelial cells, and promoted angiogenesis in the zebrafish embryo and chorioallantoic membrane of the chick embryo. In addition, there were lower levels of myristic acid and lauric acid, and higher contents of 20-hydroxyeicosatetraenoic acid (20-HETE) in CYP4Z1-expressing T47D cells compared with vector control. CYP4Z1 overexpression significantly increased tumor weight and microvessel density by 2.6-fold and 1.9-fold in human tumor xenograft models, respectively. Moreover, CYP4Z1 transfection increased the phosphorylation of ERK1/2 and PI3K/Akt, while PI3K or ERK inhibitors and siRNA silencing reversed CYP4Z1-mediated changes in VEGF-A and TIMP-2 expression. Conversely, HET0016, an inhibitor of the CYP4 family, potently inhibited the tumor-induced angiogenesis with associated changes in the intracellular levels of myristic acid, lauric acid and 20-HETE. Collectively, these data suggest that increased CYP4Z1 expression promotes tumor angiogenesis and growth in breast cancer partly via PI3K/Akt and ERK1/2 activation.

Dopamine induces growth inhibition and vascular normalization through reprogramming M2-polarized macrophages in rat C6 glioma

“…The conditioned media were collected as previously described (Yu et al, 2012). Briefly, the confluent pMΦs and RAW264.7 cells treated with 20 ng/ml of mouse recombinant IL-4 and IL-13 for 12 h were incubated with DA (10-40 μM) or vehicle for another 24 h. In another experiment, the two M2 macrophages as described above were pretreated with or without butaclamol (Buta, 50 μM), eticlopride (Eti, 50 μM) for 1 h, or DR2-siRNA, scram-siRNA for 24 h, and then incubated with DA (40 μM) for 24 h. The conditioned media were harvested, and then subjected to centrifugation through an Amicon Ultra-4 filter to remove any traces of DA, Buta and Eti.…”

Tumor Microenvironment Heterogeneity: Challenges and Opportunities

“…It was found that CYP4Z1 overexpression promotes tumor angiogenesis and growth in breast cancer. Increased CYP4Z1 expression promotes tumor angiogenesis and growth in breast cancer, partly via PI3K/Akt and ERK1/2 activation (Yu et al, 2012). ECM stiffness enhances cancer invasion and aggressive tumor cell behavior through TGF–β signaling and immune cell infiltration (Acerbi et al, 2015; Chaffer et al).…”

Vascular actions of 20-HETE

“…In an in vivo , tumor xenograft model, human non-small cell lung cancer cells expressing CYP4A11 showed an increase in tumor volume, microvessel density, and lung metastasis [116]. Another recent study indicated that breast cancer cells with overexpression of the novel CYP4 family member CYP4Z1 generated increased 20-HETE levels [117]. Media from CYP4Z1-expressing T47D and BT-474 breast cancer cells contained angiogenic factors: this media promoted proliferation, migration, and tube formation of human umbilical vein endothelial cells and promoted angiogenesis in zebrafish and chick embryos [116].…”