Vascular endothelial growth factor-A is widely regarded as the principal stimulator of angiogenesis required for tumour growth. VEGF is generated as multiple isoforms of two families, the pro-angiogenic family generated by proximal splice site selection in the terminal exon, termed VEGF xxx , and the anti-angiogenic family formed by distal splice site selection in the terminal exon, termed VEGF xxx b, where xxx is the amino acid number. The most studied isoforms, VEGF 165 and VEGF 165 b have been shown to be present in tumour and normal tissues respectively. VEGF 165 b has been shown to inhibit VEGF- and hypoxia-induced angiogenesis, and VEGF-induced cell migration and proliferation in vitro . Here we show that overexpression of VEGF 165 b by tumour cells inhibits the growth of prostate carcinoma, Ewing's sarcoma and renal cell carcinoma in xenografted mouse tumour models. Moreover, VEGF 165 b overexpression inhibited tumour cell-mediated migration and proliferation of endothelial cells. These data show that overexpression of VEGF 165 b can inhibit growth of multiple tumour types in vivo indicating that VEGF 165 b has potential as an anti-angiogenic, anti-tumour strategy in a number of different tumour types, either by control of VEGF 165 b expression by regulation of splicing, overexpression of VEGF 165 b, or therapeutic delivery of VEGF 165 b to tumours.

Angiogenesis plays an essential role in tumor growth and metastasis and is a promising therapeutic target for cancer. Vascular endothelial growth factor (VEGF) is a key regulator in vasculogenesis as well as in angiogenesis.TC71human Ewing's sarcoma cells overexpressVEGF, with a shift in isoform production from membrane-bound VEGF 189 to the more soluble VEGF 165 . Transfection of TC71cells with a vector-based VEGF targeted small interfering RNA expression system (VEGFsi) inhibited VEGF 165 expression by 80% and VEGF 165 protein production by 98%, with no alteration in VEGF 189 expression. Human microvascular endothelial cell proliferation and migration induced by conditioned medium from VEGFsi-transfected TC71 cells was significantly less than that induced by conditioned medium fromTC71cells and control vector-transfected TC71cells. Furthermore, after s.c. injection into athymic nu/nu mice, the tumor growth of VEGFsi-expressingTC71 cells was significantly less than that of parental or control vector-transfected cells.Vessel density as assessed by CD31 immunohistochemical analysis and VEGF 165 expression as assessed by Northern blotting were also decreased. Intratumor gene therapy with polyethylenimine/VEGFsi also resulted in tumor growth suppression.When inoculated into the tibias of nude mice,VEGFsiexpressingTC71cells induced osteolytic bone lesions that were less severe than those induced by control groups. These data suggest that targeting VEGF 165 may provide a therapeutic option for Ewing's sarcoma.

20-Hydroxyeicosatetraenoic acids (20-HETE), a biologically active cytochrome P-450 (CYP) metabolite of arachidonic acid in the rat kidney, can be catalyzed by CYP4A isoforms including CYP4A1, CYP4A2, and CYP4A3. To determine the contribution of CYP4A isoforms to renal 20-HETE synthesis, specific antisense oligonucleotides (ODNs) were developed, and their specificity was examined in vitro in Sf9 cells expressing CYP4A isoforms and in vivo in Sprague-Dawley rats. Administration of CYP4A2 antisense ODNs (167 nmol ⋅ kg body wt−1 ⋅ day−1iv for 5 days) decreased vascular 20-HETE synthesis by 48% with no effect on tubular synthesis, whereas administration of CYP4A1 antisense ODNs inhibited vascular and tubular 20-HETE synthesis by 52 and 40%, respectively. RT-PCR of microdissected renal microvessel RNA indicated the presence of CYP4A1, CYP4A2, and CYP4A3 mRNAs, and a CYP4A1-immunoreactive protein was detected by Western analysis of microvessel homogenates. Blood pressure measurements revealed a reduction of 17 ± 6 and 16 ± 4 mmHg in groups receiving CYP4A1 and CYP4A2 antisense ODNs, respectively. These studies implicate CYP4A1 as a major 20-HETE synthesizing activity in the rat kidney and further document the feasibility of using antisense ODNs to specifically inhibit 20-HETE synthesis and thereby investigate its role in the regulation of renal function and blood pressure.

BACKGROUND: The most important indicator for urologic surgeons at The Johns Hopkins Hospital to have a patient undergo cystoscopy is a cytologic diagnosis of high‐grade urothelial carcinoma. The template was designed to standardize diagnostic categories so clinicians can manage their patients uniformly. The template was based in part on the Bethesda System for cervical cytology. METHODS: According to the template, reactive/inflammatory changes were included in the negative group (no urothelial atypia or malignancy identified). The category atypical urothelial cells of undetermined significance (AUC‐US) was akin to atypical squamous cells of undetermined significance (ASC‐US), as was the category of atypical urothelial cells, favor high‐grade carcinoma (AUC‐H). The categories high‐grade urothelial carcinoma (HGUC) and low‐grade urothelial carcinoma also were added. RESULTS: The Pathology Data System at the Johns Hopkins Hospital was searched for cases that met the following criteria over a period from July 1, 2007 to June 30, 2009: all cytologic specimens from the urinary tract and all surgical specimens with a diagnosis of HGUC, regardless of invasion status. All cytologic specimens were then matched with biopsies during the same period, and all surgical specimens from patients who had a cytologic diagnosis of AUC‐US or AUC‐H were retrieved for 18 months after the end of the 2‐year study period. Greater than 50% of patients who had biopsy‐confirmed HGUC had a preceding cytologic diagnosis of AUC‐H or HGUC. When patients with AUC‐US were added to the analysis, 80% of patients with HGUC had at least 1 abnormal urinary cytology result. Of those patients who had a diagnosis of AUC‐H, 38% had urothelial cancer discovered at biopsy compared with only 10% of those with an AUC‐US diagnosis. CONCLUSIONS: The authors concluded that their template is effective in targeting those patients who need to undergo cystoscopy. Cancer (Cancer Cytopathol) 2013;121:15–20 © 2012 American Cancer Society.

Upon T cell activation, IkappaB kinases (IKKs) are transiently recruited to the plasma membrane-associated lipid raft microdomains for activation of NF-kappaB in promoting T cell proliferation. Retroviral Tax proteins from human T cell leukemia virus type 1 and type 2 (HTLV-1 and -2) are capable of activating IKK, yet only HTLV-1 infection causes T cell leukemia, which correlates with persistent activation of NF-kappaB induced by Tax1. Here, we show that the Tax proteins exhibit differential modes of IKK activation. The subunits of IKK are constitutively present in lipid rafts in activated forms in HTLV-1-infected T cells that express Tax. Disruption of lipid rafts impairs IkappaB kinase activation by Tax1. We also show that the cytoplasmic Tax1 protein persistently resides in the Golgi-associated lipid raft microdomains. Tax1 directs lipid raft translocation of IKK through selective interaction with IKKgamma and accordingly, depletion of IKKgamma impairs Tax1-directed lipid raft recruitment of IKKalpha and IKKbeta. In contrast, Tax2 activates NF-kappaB in a manner independent of lipid raft recruitment of IKK. These findings indicate that Tax1 actively recruits IKK to the lipid raft microdomains for persistent activation of NF-kappaB, thereby contributing to HTLV-1 oncogenesis.