TGF alpha L3-SEB fusion protein as an anticancer against ovarian cancer

TGF alpha L3-SEB is a new synthetic fusion protein produced by the combination of the third loop of transforming growth factor with staphylococcal enterotoxin type B. In the current study, the anti-tumor effects of TGF alpha L3-SEB were evaluated against SKOV3 cells, which highly expressed the epidermal growth factor receptor (EGFR). Our findings showed that incubation of SKOV3 cells with 75, 100 and 150 mu g/ml of TGF alpha L3-SEB significantly reduces the proliferation rate in a concentration-dependent manner (P < 0.05) and its IC50 value was 110 mu g/ml. Caspase-3 activity was increased from 100 for control cells to 109, 144, and 169 for 75, 100 and 150 mu g/ml of TGF alpha L3-SEB treatment, respectively. Caspase-9 activity and bax/bcl-2 ratio were also confirmed the apoptosis induction ability of TGF alpha L3-SEB (P < 0.001). Flow cytometry examination also showed that apoptosis was induced and the number of apoptotic cells was increased from 8.2 in un-treated cells to 20.9, 50, and 90 in response to 75, 100 and 150 mu g/ml of TGF alpha L3-SEB fusion protein in a concentration-dependent manner (P < 0.05). The mRNA expression level of VEGF was also reduced to 0.89, 0.69, and 0.60, respectively in response to 75, 100 and 150 mu g/ml of TGF alpha L3-SEB fusion protein exposure, respectively (P < 0.5). In summary, the findings of our study uncovered that TGF alpha L3-SEB fusion protein induced apoptosis and reduced angiogenesis in SKOV3 ovarian cancer cells in a concentration-dependent manner. This protein has the potential to act against EGFR expressing malignant cells to serve as a pro-apoptotic and angiogenesis blocker agents; however, further studies are needed to confirm its ability

Growth-inhibitory effects of TGFαL3-SEB chimeric protein on colon cancer cell line

Background: TGFαL3-SEB chimeric protein is a synthetic protein, which is produced by combining the third loop (L3) of transforming growth factor-α (TGF-α) with staphylococcal enterotoxin type B. To the best of our knowledge, anti-cancer activity of this chimeric protein against colon cancer that overexpresses epidermal growth factor receptor (EGFR) has not yet been studied. Thus, in the present study, the anti-tumor effects of TGFαL3-SEB chimeric protein on HT-29 colon cancer cells were evaluated. Materials and methods: The TGFαL3-SEB chimeric protein was previously designed and cloned in Escherichia coli (E. coli) 1,2. The level of expression and the purity of this novel protein were examined for further analysis. For this purpose, the cells were treated with different concentrations (25, 50 and 75 μg/ml) of TGFαL3-SEB and then the proliferation was detected using the MTT assay. The apoptosis-inducing potential of TGFαL3-SEB in HT-29 and HEK-293 cells was evaluated by flow cytometry using Annexin V/PI double staining method; in addition, bax/bcl2 mRNA ratio, caspase-3 and caspase-9 activity were also assessed. Results: In the present study, TGFαL3-SEB chimeric protein was produced in E. coli. After effective purification, its growth inhibitory effect was evaluated. Our results indicated that the incubation of HT-29 colon cancer cell with 25, 50 and 75 μg/ml of TGFαL3-SEB for 24 h leads to significant reduction of proliferation in a dose-dependent manner (P < 0.05). Further analysis indicated that exposure of EGFR expressing HT-29 cells to TGFαL3-SEB leads to significant increase of the caspase-3 and caspase-9 activity in a concentration-dependent manner (P < 0.05). Bax/bcl-2 ratio also confirmed that TGFαL3-SEB has the pro-apoptotic effect. Flow cytometry analysis of TGFαL3-SEB treated cells showed that in addition to apoptotic cells, necrotic cells were also increased significantly at the concentration of 25, 50 and 75 μg/ml (P < 0.05). Conclusion: In conclusion, our results demonstrated that TGFαL3-SEB chimeric protein induced cell death through both mechanisms of apoptosis and necrosis in HT-29 colon cancer cells. This paper has highlighted that TGFαL3-SEB has the potential to target EGFR expressing cancer cell. © 2018 The Author

Application pratique et intérêt des recommandations de prise en charge des syncopes : évaluation de pratiques professionnelles

AIMS: To assess the practical implementation of international guidelines and their impact on syncope management in a 500-bed general hospital. PATIENTS AND METHODS: Three groups of 63 consecutive patients admitted for syncope to the emergency care unit (ECU) were studied: group 1, before the guidelines delivered to the practitioners, group 2 immediately after the diffusion of guidelines and group 3, one year later. The study evaluates the mean duration of stay (MDS) and the relevance of the diagnostic strategy. RESULTS: In group 1 compared to group 2, MDS were respectively 6.8±5.5 and 5.4±2.8 days (P=0.07) and the unexplained syncope number respectively 22% and 24% (P=0.8). The search of orthostatic hypotension became more systematic (13% versus 86% in group 1 and 2 respectively, P<0.001). The agreement (kappa coefficient) between initial and final diagnostic increased in 0.34 to 0.44. One year later MDS in group 3 was 7.1±4.7 days (P=0.8 versus group 1 and P=0.015 versus group 2) with only 6.3% systematic search for orthostatic hypotension (P<0.001). CONCLUSIONS: Guidelines optimize the syncope management in the ECU and the agreement between the emergency and discharge diagnostic without change of unexplained syncope and. MDS tend to be shorter when guidelines are actively implemented. Nevertheless, the positive impact of guidelines implementation is of limited duration