| TNF related apoptosis inducing ligand,TRAIL was newly discovered member of TNF surperfamlily.Five distinct cell surface TRAIL receptors (TRAILs) have been identified.Death receptors TRAIL-R1(DR4), TRAIL -R2(DR5) contain a cytoplasmic death domain and signal for cell death and induce cell death combined with TRAIL whereas the other three (TRAIL-R3,TRAIL-R4 and OPG) serve as decoy receptors to block TRAIL-mediated apoptosis.Targeted on death receptors tumor bilogical therapy is good candidate for cancer therapy .people have constrcted lots of TRAIL recombinant and studied many about them. In the present study,administration of soluble recombinant TRAILin experimental animals induces significant tumor regression without system toxicity. However,the ability of soluble human TRAIL to induce apoptosis of normal hepatocytes in vitro has raised question regarding the potential usefulness of this agent in cancer therapy.now study aims is to find the agent of high killing tumor and lower cytotoxicity.Ichikawa generated novel anti-DR5 monoclonal antibody TRA-8 and testing TRA-8 can not induce apoptosis of normal hepatocellular cell death but induCed apoptosis of primary hepatocellular carcinoma cells and an established liver cancer cell line .In present, Tumoricidal activity of anti-human DR monoclonal antibody on glioma cells U343,U138,U373 is not reported .so we will study on the cytotoxicity activity and mechanism of anti-human DR monoclonal antibody on glioma,giving contribution for clinical cancer therapy. The ability of anti-DR4McAb/FUM1.4,anti-DR5McAb/FUM1.5 innduce apoptosis of glioma cell lines was studied as following two steps: Part one FMU1.4/FMU1.5 induCed U343,U138,U373 cells lines apoptosis1 Expression of DR4,DR5 on glioma cell lines DR4,DR5 expression is analysed,protein were tested through FACS; mRNA were tested bthrough RT-PCR and distribution were tested by immunocytochemistry.The results showed the expression of DR4/DR5 is different,all the glioma cells expressed DR5 and DR4 on the cell surface and DR5 expression on U343,U138,U373 was 86.5%,22% 2.61% respectively;DR4 expression was very low on U373 only 0.11%, U343, U138 was 53%,2.08% respectively.DR5/DR4 mRNA were different , DR5 mRNA is tested in U343,U138,U373,DR4 is not tested in U373. Conclusion DR4/DR5 protein expression is correlated with mRNA expression. By immunocytochemistry DR5 was shown to be present in U343U138U373 and DR5 appeared to be located in intracellular perinuclear compartment as has been shown previously.2 FUM1.4/FUM1.5 killing Human Glioma Cell Lines Three glioma cell lines U343,U138,U373 were exposed to FUM1.4,FUM1.5 for 4h , cytotoxicity was compared by MTT assay,FACS,and resulte indicated U343 exhibit sensitivity to FUM1.5-mediated cytotoxicity in a concentration-dependent mannerï¼›however ,the other two cell lines is not:U373 was resistant to FUM1.5,U138 was partly sensitivity compared to U373.U343 was partly sensitivity to FUM1.4,the other two cell lines were resistant to FUM1.4. In sum,U343 was sensitivity to FUM1.5. cytotoxicity of FUM1.5 was stronger than FUM1.4.3 Morphological Observation of Apoptotic Glioma Cell LinesU343 were received after FUM1.5 40ug/mL added to cells: observation under light microscope and transmission electron microscope, the apoptotic cells occur irregular morphology,nucleus condensation chromatin margin or apoptotic bodied formation,but the mitochondra basically remain integrity.There are few apoptotic glioma cells in control groups.4 DNA Fragmentation Qualitative and Quantitative AnalysisU343,U138,U373 are cultured with 40ug/mL FUM1.5 and the suspended cells are collected after 4h culture.The agarose gel electrophoresis diphenylamin colorimetry are used to detect DNA fragmentation .It is indicated that the genomic DNA occurs "ladder "fragmentation in U343 cultured by FUM1.5/FUM1.4,but genomic DNA from other groups did not display any visible fragmentation bands .The percentage of DNA fragmentation was analysed by FACS and showe...
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