MEK1 Inhibitor Combined with Irradiation Reduces Migration of Breast Cancer Cells Including miR-221 and ZEB1 EMT Marker Expression.

The miR-221 expression is dependent on the oncogenic RAS-RAF-MEK pathway activation and influences epithelial-to-mesenchymal transition (EMT). The Cancer Genome Atlas (TCGA) database analysis showed high gene significance for ZEB1 with EMT module analysis and miR-221 overexpression within the triple-negative breast cancer (TNBC) and HER2+ subgroups when compared to luminal A/B subgroups. EMT marker expression analysis after MEK1 (TAK-733) inhibitor treatment and irradiation was combined with miR-221 and ZEB1 expression analysis. The interaction of miR-221 overexpression with irradiation and its influence on migration, proliferation, colony formation and subsequent EMT target activation were investigated. The results revealed that MEK1 inhibitor treatment combined with irradiation could decrease the migratory potential of breast cancer cells including reduction of miR-221 and corresponding downstream ZEB1 (EMT) marker expression. The clonogenic survival assays revealed that miR-221 overexpressing SKBR3 cells were more radioresistant when compared to the control. Remarkably, the effect of miR-221 overexpression on migration in highly proliferative and highly HER2-positive SKBR3 cells remained constant even upon 8 Gy irradiation. Further, in naturally miR-221-overexpressing MDA-MB-231 cells, the proliferation and migration significantly decrease after miR-221 knockdown. This leads to the assumption that radiation alone is not reducing migration capacity of miR-221-overexpressing cells and that additional factors play an important role in this context. The miR-221/ZEB1 activity is efficiently targeted upon MEK1 inhibitor (TAK-733) treatment and when combined with irradiation treatment, significant reduction in migration of breast cancer cells was shown.

Voluntary STD testing and treatment program at a metropolitan correctional facility: evaluation of test acceptability and associated risk factors.

Few studies have addressed challenges of diagnosis and treatment of sexually transmitted diseases (STDs) within correctional facilities. Initiatives that screen all inmates can be cost-prohibitive, while symptom-based screening undoubtedly fails to recognize significant numbers of asymptomatically infected persons. This study discusses a voluntary STD screening and treatment program developed at the Douglas County (Nebraska) Department of Corrections where student volunteers interviewed, screened, and educated 456 inmates. Inmate urine samples and interview responses about risk behaviors and motivators for participation in the screening program were analyzed. The results support the ongoing project method to screen and treat inmates in the community correctional facility. Risk factor analysis suggests that targeted testing and treatment efforts may have a role in providing cost-effective care for STD among the incarcerated population.

Analysis of the maturation process of dendritic cells deficient for TNF and lymphotoxin-alpha reveals an essential role for TNF.

Dendritic cells (DCs) generated from bone marrow (BM) precursor cells of C57BL/6 (B6.WT) mice and cultured in the presence of granulocyte macrophage-colony stimulating factor differentiate to mature BM-DCs spontaneously. These mature DCs are characterized by high levels of major histocompatibility complex (MHC) class II, CD40, and CD86 on their surface. To analyze the involvement of tumor necrosis factor (TNF) and the related cytokine lymphotoxin (LT)alpha in DC maturation, we studied the development of DCs from the BM of B6.TNF(-/-), B6.LTalpha(-/-), and B6.TNF/LTalpha(-/-) mice and compared it to B6.WT mice. Although the development of BM precursor cells to the level of immature DCs (CD11c(+), MHC class II(low), CD40(low), and CD86(low)) was equivalent in all genotypes, B6.TNF(-/-) and B6.TNF/LTalpha(-/-) cells showed an impaired capacity to differentiate to mature DCs. In contrast, mature BM-DCs generated from LTalpha-negative, immature DCs developed like B6.WT cells. Further studies revealed that once matured, the phenotype of all tested genotypes was comparable. They expressed high levels of CD40 and CD86, were exclusively positive for the chemokine receptor (CCR)7 but negative for CCR5 and CCR2, and were able to enter the paracortex of draining lymph nodes. The limited maturation of TNF-deficient BM-DCs could be restored by mixing TNF-negative with TNF-positive Ly5.1 BM cells, and maturation of B6.WT DCs could be blocked with an anti-TNF monoclonal antibody. The substitution of B6.TNF(-/-) BM cells with recombinant TNF revealed promotion or suppression of BM-DC maturation depending on the point of time of TNF addition.