MicroRNA-204/CREB5 axis regulates vasculogenic mimicry in breast cancer cells.

BACKGROUND: Vasculogenic mimicry (VM) is characterized by formation of three-dimensional (3D) channels-like structures by tumor cells, supplying the nutrients needed for tumor growth. VM is stimulated by hypoxic tumor microenvironment, and it has been associated with increased metastasis and clinical poor outcome in cancer patients. cAMP responsive element (CRE)-binding protein 5 (CREB5) is a hypoxia-activated transcription factor involved in tumorigenesis. However, CREB5 functions in VM and if its regulated by microRNAs remains unknown in breast cancer. OBJECTIVE: We aim to study the functional relationships between VM, CREB5 and microRNA-204-5p (miR-204) in breast cancer cells. METHODS: CREB5 expression was evaluated by mining the public databases, and using RT-qPCR and Western blot assays. CREB5 expression was silenced using short-hairpin RNAs in MDA-MB-231 and MCF-7 breast cancer cells. VM formation was analyzed using matrigel-based cultures in hypoxic conditions. MiR-204 expression was restored in cancer cells by transfection of RNA mimics. Luciferase reporter assays were performed to evaluate the binding of miR-204 to 3'UTR of CREB5. RESULTS: Our data showed that CREB5 mRNA expression was upregulated in a set of breast cancer cell lines and clinical tumors, and it was positively associated with poor prognosis in lymph nodes positive and grade 3 basal breast cancer patients. Silencing of CREB5 impaired the hypoxia-induced formation of 3D channels-like structures representative of the early stages of VM in MDA-MB-231 cells. In contrast, VM formation was not observed in MCF-7 cells. Interestingly, we found that CREB5 expression was negatively regulated by miR-204 mimics in breast cancer cells. Functional analysis confirmed that miR-204 binds to CREB5 3'-UTR indicating that it's an ulterior effector. CONCLUSIONS: Our findings suggested that CREB5 could be a potential biomarker of disease progression in basal subtype of breast cancer, and that perturbations of the miR-204/CREB5 axis plays an important role in VM development in breast cancer cells.

Reduction of dopaminergic transmission in the globus pallidus increases anxiety-like behavior without altering motor activity.

The globus pallidus (GP) plays an important role in the flow of information between input and output structures of the basal ganglia (BG) circuit. In addition to participating in motor control, the GP may also be involved in cognitive and emotional functions related to the symptoms of patients with Parkinson's disease (PD). Since the GP receives dopaminergic innervation from the substantia nigra pars compacta (SNc), it is important to determine whether a local dopamine (DA) deficit in the GP is related not only to motor but also to the cognitive and emotional alterations of PD. The aim of this study was to examine the effects of lesions in the GP (induced by 6-OHDA) on anxiety, depression and ambulation in rats. Such lesions are known to reduce dopaminergic innervation in this brain structure. Additionally, the effect on DA receptors in the GP was tested by local administration of the dopamine agonist PD168,077, antagonist haloperidol and psychostimulant amphetamine. Experimental anxiety was evaluated with the elevated plus maze (EPM), burying behavior test (BBT) and social interaction test, while depressive-like behavior was assessed with the sucrose preference test. Rats with unilateral and bilateral lesions showed a higher level of anxiety than intact animals in both the EPM and BBT, an effect also obtained after intrapallidal injection of haloperidol. The administration of methamphetamine or PD-168.077 caused the opposite effect. The dopaminergic lesions in the GP did not affect sucrose preference, social interaction or ambulation. These results show that dopamine in the GP, acting through D2 or D4 receptors, may be involved in the manifestation of anxiety, a non-motor symptom of PD that often appears before motor symptoms.

In vitro anti-Vibrio cholerae activity of essential oil from Lepechinia caulescens.

Lepechinia caulescens is a plant employed by the Purepecha (a Mexican ethnic group) to treat infectious gastrointestinal ailments. The essential oil of this species was active against some strains of Vibrio cholerae with 4 microl/ml MIC and 6 microl/ml MBC. The major components of the oil found by GC-MS were borneol, camphor and trans-caryophyllene.