ABSTRACT
Objective:To study the cytotoxic effects on three glioma cell lines U343, U138, U373 induced by anti-human DR5/DR4 monoclonal antibodies(FMU1.5/FMU1.4) and the underlying mechanism.Methods:Expression of DR4/DR5 was quantitated by flow cytometry and DR/4DR5 mRNA detected by RT-PCR. Cytotoxicity exerted by FMU1.4/FMU1.5 on three cell lines was measured by MTT colorimetry and the induced apoptosis was determined by agarose gel electrophoresis, DNA ploidy analysis was studied by flow cytometry.Results:The expression of DR5 on U343 cells was higher and the expression of DR4 on U373 cells was lower. Cell line U343 was sensitive to FMU1.5 and in a dose dependent manner, but it was partially sensitive to FMU1.4; Cell line U138 was partially sensitive to FMU1.5 and resistant to FMU1.4; Cell line U373 was insensitive to two antibodies.Conclusion:Apoptosis induced by monoclonal antibodie FMU1.4/FMU1.5 vary among three cell lines. The underlying mechanism may be relevant to DR4/DR5 expression,the release of cytochrome C and FLIP.
ABSTRACT
Objective: To investigate the inhibitory effects and mechanism of a novel anti-human DR5 ( death receptor 5 of TRAIL) monoclonal antibody to glioma cell lines U343. Methods: DR5 protein was tested quantitative through FCM: DR5 mRNA was observed through RT-PCR and distribution was tested by immunocytochemistry. Inhibitory effects and inducing apoptosis of anti-human DR5 monoclonal antibody to U343 were analysed by MTT, DNA Ladder, FCM. Results: The expression of death receptor 5 ( DR5 ) was certificated in U343 , DR5 appeared to be located in intracellular perinucle-ar compartment. Inhibitory effects of anti-human DR5 monoclonal antibody on U343 were achieved by 3 ?g/ml at 4 hours and the mechanism was associated with apoptosis. Conclusion: Apoptosis of glioma cell lines U343 can be induced by anti-human DR5 monoclonal antibody, and targeted on DR will provide new way to treating cancer.
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Objective:To study the bioactivity of thymosin ?16 (T?16) in vitro and in vivo.Methods:Recombinant His-SUMO-T?16 was constructed and transformed into E.coli BL21(DE3) for induced expression.The product was treated by ultrasonication,ion-exchange chromatography and metal chelation chromatography respectively for purification.The fusion protein was cut by His-SUMO protease and then further purified by metal chelation chromatography and Superdex 30 gel chromatography.Results:Recombinant fusion protein His-SUMO-T?16 was soluble,whose specific activity was 5.3?105 U/mg.It could promote the proliferation of BALB/c 3T3 cells,rabbit corneal cells,and chicken embryo chorion vessels in vitro,and both the proliferation and migration of vascular endothelial cells in vitro were enhanced,and rabbit skin healing of alkali burns in vivo was accelerated.Conclusion:E.coli expressing vector of recombinant His-SUMO-T?16 fusion protein is constructed successfully,and recombinant protein T?16 has significant repairing effects.The study established a good foundation for further industrialization of T?16.
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Objective:To study the effects of IL-2R? mimic peptide(CP) on skin allograft rejection in mice.Methods:Mouse skin allograft model was employed in all groups.The bioactivities were determined by lymphocyte proliferation,one-way MLR,assay of T lymphocyte subset and the level of cytokine in splenic cell culture supernatant.Results:CP inhibited lymphocyte proliferation;decreased the number of CD4+T lymphocytes in spleen and the value of CD4+/CD8+;down-regulated the level of IL-2 and IFN in splenic cell culture supernatant,as well as prolonged the mean survival times(MST) of skin allografts.The combination of CP and CsA showed cooperativities.Conclusion:CP,as the IL-2R? mimic peptide has the suppressive activities,which can efficiently inhibit skin allograft rejection in mice.