Combined effect of the pro-apoptotic rhTRAIL protein and HSV-1 virus in head and neck cancer cell lines.

Knowledge on the molecular and clinical characteristics of head and neck squamous cell carcinoma (HNSCC) is vast. However, an effective therapy that increases the life expectancy of these patients, with a 5-year overall survival of 50%, is still unknown. Here we evaluated the combined effect of the pro-apoptotic protein rhTRAIL with the replication-competent wild-type HSV-1 virus in head and neck cancer cell lines. We observed a difference in the modulation profile of proteins related to apoptotic pathways in the studied cell lines. The HCB289 exhibited caspase-9 activation in the presence of the HSV-1 virus, while the UD-SCC-2 exhibited caspase-8 activation in the presence of rhTRAIL. Both cell lines exhibited PARP activation by combining rhTRAIL and HSV-1 virus treatment. Flow cytometry analysis exhibited greater induction of late apoptosis for the HCB289 and UD-SCC-2 after the combination treatment of the HSV-1 and rhTRAIL. However, the UD-SCC-2 also presented induction of late apoptosis by the presence of rhTRAIL in monotherapy. These data suggest an enhancement of the effect of the combination treatment of the rhTRAIL and the HSV-1 on reducing viability and induction of cell death.

Risk factors for depression in adults: NR3C1 DNA methylation and lifestyle association.

OBJECTIVE: The aim of this study was to verify determinant factors for depression and analyze the relationship between possible changes in HPA axis and depression, in this case NR3C1 DNA methylation and serum cortisol levels. METHODS: 349 adult volunteers were recruited to evaluate depression, socio-demographic, economic and lifestyle factors, serum cortisol levels and NR3C1 DNA methylation by pyrosequencing. Depression determinant factors were investigated using a Poisson regression model with robust variance (p < 0.05). RESULTS: Poisson regression with robust variance adjusted by gender, tobacco use, self-perceived stress, leisure activity, suicidal ideation, low cortisol levels and NR3C1 DNA methylation was performed and predicted risk factors for depression. Furthermore, depressive volunteers showed a significant increase in NR3C1 DNA methylation when compared to healthy volunteers. CONCLUSIONS: This findings provide a basis for understanding the role of HPA axis in depression, especially its regulation by NR3C1 DNA methylation. Furthermore, it emphasizes the stressful lifestyle risk factors (female, tobacco uso, self perceived stress, leisure activities absence and suicidal ideation) that can contribute to future research and the search for public health policies to improve quality of live, mental and general health.

Construction and characterization of a new TRAIL soluble form, active at picomolar concentrations.

Apoptosis induction has emerged as a treatment option for anticancer therapy. Tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), a type II transmembrane protein, is a potent and specific pro-apoptotic protein ligand, which activates the extrinsic apoptosis pathway of the cell death receptors. Here we describe the construction and characterization of a new soluble TRAIL, sfTRAIL, stabilized with the trimerization Foldon domain from the Fibritin protein of the bacteriophage T4. Supernatants of 0.22 µM-filtered supernatants were produced in Vero-transduced cells with HSV1-derived viral amplicon vectors. Experiments were undertaken in two known TRAIL-sensitive (U373 and MDA.MB.231) and two TRAIL-resistant (MCF7 and A549) cell lines, to determine (i) whether the sfTRAIL protein is synthetized and, (ii) whether sfTRAIL could induce receptor-mediated apoptosis. Our results showed that sfTRAIL was able to induce apoptosis at concentrations as low as 1899.29 pg/mL (27.71 pM), independently of caspase-9 activation, and reduction in cell viability at 998.73 fM.