Preferential down-regulation of phospholipase C- β in Ewing's sarcoma cells transfected with antisense EWS-Fli-1

EWS-Fli-1, a fusion gene found in Ewing's sarcoma and primitive neuro-ectodermal tumour (PNET), encodes a transcriptional activator and promotes cellular transformation. We have made stable Ewing's sarcoma cells expressing antisense EWS-Fli-1 transcripts by transfecting the antisense EWS-Fli-1 expression plasmid. These cells showed partial loss of endogenous EWS-Fli-1 proteins and suppression of the cell growth. To elucidate the molecular mechanisms underlying the growth inhibition, we examined the changes of signal transducing proteins by immunoblot analysis in Ewing's sarcoma cells stably expressing antisense EWS-Fli-1 transcripts. Western blotting of the cell proteins revealed that expressions of phospholipase Cβ2 and β3 (PLCβ2, PLCβ3), and also protein kinase C α and β (PKCα, β) were significantly reduced by transfecting with antisense EWS-Fli-1. The inositol phosphates production by bradykinin (BK), but not platelet-derived growth factor (PDGF), was suppressed in these cells. These results suggest that the PLCβ2 and PLCβ3 may play a role in tumour proliferation in Ewing's sarcoma cells. © 2000 Cancer Research Campaig

White Paper: functionality and efficacy of wrist protectors in snowboarding—towards a harmonized international standard

The wrist is the most frequently injured body region among snowboarders. Studies have shown that the risk of sustaining a wrist injury can be reduced by wearing wrist protection. Currently, there are a wide variety of wrist protection products for snowboarding on the market that offer a range of protective features. However, there are no minimum performance standards for snowboarding wrist protectors worldwide. The International Society for Skiing Safety convened a task force to develop a White Paper to evaluate the importance and necessity of a minimum performance for all wrist protectors used in snowboarding. The White Paper outlines the need for a general framework for a harmonized international standard and reviews the existing evidence. Therefore, this White Paper may serve as a common base for future discussions. The broader goal of developing and implementing such a standard is to reduce the incidence and the severity of wrist injuries in snowboarding without increasing the risk of adverse events, such as upper arm or shoulder injury. The European standard for inline skating wrist protectors (EN 14120) can serve as a starting point for efforts related to a standard for snowboard wrist protectors, but certain modifications to the standard would be required. It is hypothesized that implementation of a snowboarding wrist protector standard would result in fewer and less severe wrist injuries in the sport and could translate into more riding days for healthy snowboarders and significant health-care costs savings

Oncogenic Fusion Protein EWS/FLI1 Down-regulates Gene Expression by Both Transcriptional and Posttranscriptional Mechanisms*

Ewing family tumors are characterized by a translocation between the RNA binding protein EWS and one of five ETS transcription factors, most commonly FLI1. The fusion protein produced by the translocation has been thought to act as an aberrant transcription factor, leading to changes in gene expression and cellular transformation. In this study, we investigated the specific processes EWS/FLI1 utilizes to alter gene expression. Using both heterologous NIH 3T3 and human Ewing Family Tumor cell lines, we have demonstrated by quantitative pre-mRNA analysis that EWS/FLI1 repressed the expression of previously validated direct target genes at the level of transcript synthesis. ChIP experiments showed that EWS/FLI1 decreases the amount of Pol II at the promoter of down-regulated genes in both murine and human model systems. However, in down-regulated target genes, there was a significant disparity between the modulation of cognate mRNA and pre-mRNAs, suggesting that these genes could also be regulated at a posttranscriptional level. Confirming this, we found that EWS/FLI1 decreased the transcript half-life of insulin-like growth factor binding protein 3, a down-regulated direct target gene in human tumor-derived Ewing's sarcoma cell lines. Additionally, we have shown through reexpression experiments that full EWS/FLI1-mediated transcriptional repression requires intact EWS and ETS domains. Together these data demonstrate that EWS/FLI1 can dictate steady-state target gene expression by modulating both transcript synthesis and degradation