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Objective To evaluate the effect of bifidobacterium-mediated CTP-NPRL2 on the growth and apoptosis of nude mouse renal carcinoma.Methods Recombinant plasmid pET15b-CTP-NPRL2 was constructed and transfected into bifidobacterium by electroporation,and then the expression of fusion protein CTP-NPRL2 was verified by Western blot.The nude mouse renal carcinoma model was constructed by subcutaneous injection of renal carcinoma cells suspension.The nude mice with renal carcinoma were equally and randomly divided into the observation group and control group(8 mice in each group).The mice in the observation group were treated with bifidobacterium containing recombinant plasmid pET15b-CTP-NPRL2 through tail vein injection,and the mice in the control group were treated with normal saline.All mice were treated once a week for four weeks,and then executed for evaluating the weight of mice and bearing tumors.Finally,the apoptosis of renal carcinoma was detected by TUNEL staining.Results The mass of nude mice was(26.24±1.98) g in the observation group and(23.28±2.17) g in the control group.The mass of bearing tumors was(1.37±0.12) g in the observation group and(1.68±0.18) g in the control group,and the differences were statistically significant(P＜0.05).The TUNEL detection results showed that the apoptosis index of renal carcinoma cell in the observation group(23.27±5.14)％ was significantly higher than that in the control group(10.37±2.58)％,and the difference was statistically significant(P＜0.05).Conclusion Bifidobacterium-mediated CTP-NPRL2 has an inhibitory effect on the renal carcinoma growth of nude mouse,and significantly increases the apoptosis of renal carcinoma cells.
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Objective: To investigate the effects of fusion peptide cytoplasmic transduction peptide-oligomerization domain 1-hemagglutinin (CTP-OD1-HA) and CTP-OD2-HA on subcutaneous xenograft of leukemia K562 cells in nude mice. Methods: K562 cells were pre-treated with CTP-OD-HA (as a positive control), CTP-HA (as a negative control), PBS (as a blank control), CTP-OD1-HA and CTP-OD2-HA, respectively, then the cells were injected into nude mice subcutaneously. The growth of the subcutaneous xenograft was observed. The apoptosis of the tumor cells in subcutaneous xenograft was detected by TUNEL method. The expression levels of Bax and Bcl-2 proteins in subcutaneous xenograft were examined by immunohistochemistry. Results: The volumes of the subcutaneous xenografts in CTP-OD1-HA group and CTP-OD2-HA group were smaller than that in CTP-HA group (P < 0.05), with the change of cavitation apoptosis. The apoptotic change of tumor cells was significant in CTP-OD1-HA group and CTP-OD2-HA group by TUNEL method (P < 0.05). The expression level of apoptosis-associated protein Bax was higher in CTP-OD1-HA group and CTP-OD2-HA group than that in CTP-HA group (P < 0.05), whereas the expression level of Bcl-2 was lower (P < 0.05). Conclusion: CTP-OD1-HA and CTP-OD2-HA fusion peptide can inhibit the tumorigenicity of leukemia K562 cells in nude mice and promote the apoptosis. Copyright © 2013 by TUMOR.
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Objective To observe the effects of cytoplasmic transduction peptide (CTP)-HBcAg18-27-Tapasin induced murine bone marrow-derived dendritic cell (DC) maturation on T lymphocyte proliferation in vitro,Methods Bone marrow derived DC isolated from BALB/c mice were cultured with recombinant granulocyte-macrophage colony-stimulating factor and recombinant interleutin (IL)-4 for 5 days followed by lipopolysaccharide added to induce DC maturation.10 μg/L CTP-HBcAg18-27-Tapasin,50 μg/L CTP-HBcAg18-27-Tapasin,10 μg/L CTP-HBcAg18-27 or RPMI-1640 were added into culture medium to induce DC maturation.DC phenotypes were analyzed by flow cytometry.The level of IL-12p70 in the supernatant was detected by enzyme linked immunosorbent assay.The proliferation of.T lymphocytes was performed by using cell counting kit-8 and intracellular cytokine of proliferative T cells were analyzed by flow cytometry.The means among groups were compared using one-way ANOVA and those between two groups were compared by least significant difference test.Results DC were cultured and induced successfully.The molecules on DC surface,such as CD80,CD86 and major histocompatibility antigen-Ⅰ were upregulated by CTP-HBcAg18-27-Tapasin.IL-12p70 level induced by 50 μg/L CTP-HBcAg18-27-Tapasin was (61.12±10.25) pg/mL,which was higher than those induced by 10 μg/L CTP-HBcAg18-27-Tapasin (50.43±10.42) pg/mL,10μg/L CTP-HBcAg18-27 (40.17±8.54) pg/mL and medium control (30.51±8.03) pg/mL (F=15.85,P=0.030 and 0.037).The proliferation of T lymphocytes induced by CTP- HBcAg18-27 -Tapasin was higher than control groups.The amounts of cytotoxic T lymphocyte (CTL) induced by 50 μg/L CTP-HBcAg18-27-Tapasin [(2.05±0.41) ％] and 10 μg/L CTP-HBcAg18-27-Tapasin [(1.06 ±0.10 )％] were both significantly higher than the 10 μg/L CTP-HBcAg18-27 group [(0.45±0.11)％] and medium group [(0.09±0.02)％,F=60.22,P=0.003].Conclusions CTP HBcAg18- 27 Tapasin could promote the differentiation and maturation of DC,and enhance the ability of DC stimulating T lymphocytes proliferation and increase CTL expression effectively.