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Objective To study the effect ofRunfei Zhisou Pills (RZP) on cell apoptosis in rat model of chronic obstructive pulmonary disease (COPD) and its mechanism.Methods Totally 32 male Wistar rats were randomly divided into control group,model group,1-month RZP group,and 3-month RZP group.Rats in 1-month and 3-month RZP group,received RZP of 0.9 g/kg once daily for 1 and 3 months respectively,and rats in control and model groups received water with equal volume for 1 month.The rat COPD model was established using tobacco smoke combined with intratracheal instillation of LPS.Morphological changes of lung tissue in COPD rats were observed with HE staining under light microscope.The VEGF levels in bronchoalveolar lavage fluid (BALF) were detected by ELISA.The VEGF and VEGFR2 protein expression in lung tissues was measured by Western blotting.Cell apoptosis in lung tissues was detected with TUNEL method,and apoptosis index (AI) was counted.Results Compared with control group,the pathological changes of lung tissue in model group were obvious,the AI significantly increased,VEGF in BALF and VEGF and VEGFR2 in lung tissue decreased significantly.Compared with model group,morphological improvement of lung tissue was obvious,the AI significantly decreased,and VEGF in BALF,VEGF and VEGFR2 in lung tissue increased significantly in 1-month and 3-month RZP groups.Conclusion RZP can inhibit cell apoptosis of COPD,and the mechanism may be related with up-regulation of VEGF and VEGFR2.
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Objective:To investigate the modulating effects of anti-epidermal growth factor monoclonal antibody Cetux- imab(C225)on the ehemosensitivity and radiosensitivity in a Docetaxel-resistant human lung adenocarcinoma cell line SPC-A-1/doeetaxel.Methods:Radiosensitivity of SPC-A-1/docetaxel was determined by clone formation experiment and quantified by calculating the enhancement ratio(ER).The growth inhibition of SPC-A-1/docetaxel cell line caused by C225 or combination of C225 and Docetaxel in different orders was detected by MTT assay.The effect of C225 on cell cy- cle distribution and apoptosis was determined by flow cytometry.Results:C225 combined with radiation significantly de- creased the number of the cell clones than radiation alone;the D_0 values were 1.73 Gy for the former and 2.39 Gy for the latter,and the enhancement ratio was 1.38.C225 alone at concentration up to 1000?g/ml for 48h had neither cytotoxic nor eytostatie effect on SPC-A-1/docetaxel in vitro.C225 administration followed by Docetaxel significantly decreased the ICw of Docetaxel(85.2?g/ml vs 128.7?g/ml).Flow cytometry demonstrated that C225 exposure induced apoptosis of SPC-A-1/docetaxel cells in a time-dependent manner.The cell in the G_0/G_1 fraction increased from(43.80?4.46)% to (60.50?6.57)%(P
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Objective:To prepare and identify monoclonal antibody specifically targeting epidermal growth factor receptor(EGFR) and(or) epidermal growth factor receptor vⅢ(EGFRvⅢ),and to investigate its inhibitory effects on human hepatocellular carcinoma Huh7-EGFRvⅢ cell-and epidermal carcinoma A431 cell-implanted tumors in nude mice.Methods: BALB/c mice were immunized with 3T3 cells stably transfected with EGFRvⅢ(3T3-EGFRvⅢ).Immunized spleen cells were fused with myeloma SP2/0 cells,and anti-EGFRvⅢ monoclonal antibody positive hybridoma cells(named 9B9 antibody and 9B9 cells,respectively) were selected and identified by ELISA.The specific interaction between 9B9 antibody and EGFRvⅢ/EGFR antigen was detected by Western blotting and immunofluorescence assay.Huh7-EGFRvⅢ cell-(human hepatocellular carcinoma Huh7 cells stably transfected with EGFRvⅢ) and epidermal cell carcinoma A431 cell-bearing mouse models were established and were divided into PBS group,Cetuximab group and 9B9 antibody group.Then,anti-tumor effect of 9B9 antibody was examined and compared with those of PBS and Cetuximab.Results: A monoclonal antibody,named 9B9 antibody,was obtained by hybridoma technique and it reacted with both EGFRvⅢ antigen and EGFR antigen as detected by Western blotting and immunofluorescence.The inhibitory rates of Cetuximab and 9B9 antibody against Huh7-EGFRvⅢ cells-implanted tumors were 42% and 46%,respectively,and those against A431 cells-implanted tumors were 85% and 86%,respectively.Conclusion: 9B9 monoclonal antibody can effectively inhibit the growth of human hepatocellular carcinoma cell-and epidermal cell carcinoma cell-implanted tumors,and the effects resemble that of Cetuximab.