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Archive > Volume 21 Issue 3 > 2014,21(3):257-262. DOI:10.3872/j.issn.1007-385X.2014.03.004 Prev Next
Basic Research
Cell-penetrating peptide-fused HSP70 gene therapy for gastric cancer in combination cytokine-induced killer cells in a xenograft nude mouse model
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Abstract:
To evaluate the efficacy of cell-penetrating peptide-fused HSP70 gene therapy in combination with the use of cytokine-induced killer (CIK) cells for gastric cancer in a xenograft nude mouse model. Methods: CMV promoter-driven adenoviral vectors expressing wild-type HSP70 (AdCMV-HSP70) and HSP70 fused with a cell-penetrating peptide of 11 arginines (AdCMV-HSP70s) were constructed. In in vitro experiments, human gastric cancer SGC-7901 and GES-1 cells were infected with these two viral vectors respectively. At 48 h after infection, cell viability was assessed by MTT assays and HSP70 protein content by Western blotting. In in vivo experiments, Sgc-7901 cells were injected subcutaneously into BALB/c nude mice. Cancer cell-challenged mice were treated every 48 h for a total of five times with AdCMV-HSP70 or AdCMV-HSP70s, either each alone or in combination with a single dose of 1×107 CIK cells (tail vein injection) prepared from normal BALB/c mice. Thirty-five days after treatment, animals were sacrificed. The number and size of tumors formed were assessed. Results: HSP70 protein content after AdCMV-HSP70s infection was significantly higher than that after AdCMV-HSP70 infection in both SGC cells (360.72±20.89 ng/ml vs 121.01±15.94 ng/ml, P<0.05) and GES-1 cells (188.62±10.82 ng/ml vs 135.00±13.96 ng/ml, P<0.05). Fusion of 11 arginine peptide into HSP70 led to significant inhibition of gastric cancer cell proliferation; at low multiplicity of infection (MOI) of 100 pfu/ml, the cell viability was (66.33±4.33)% in AdCMV-HSP70s-infected SGC-7901 cells, significantly lower than that (101.33±7.64%) in AdCMV-HSP70-infected SGC-7901 cells (P<0.05). CIK cell administration resulted in immune reconstruction in nude mice carrying xenograft gastric cancer cells. Adenoviral delivery of HSP70 enhanced infiltration of CD3+ T cells into tumor tissues and induce antitumor immune response. CIK cells in combination with AdCMV-HSP70s showed a significantly higher tumor inhibition rate (66.5%) than CIK cells in combination with AdCMV-HSP70 (43.6%) in nude mince challenged with gastric cancer cells (P<0.05). Conclusion: In nude mice carrying human gastric cancer cells, HSP70 may not only inhibit gastric cancer cell proliferation but also induce antitumor immune response. These effects can be enhanced by fusion of a cell-penetrating peptide with the HSP70 molecule and implantation of cytokine-induced killer cells.
Keywords:
Project Supported:
Supported by the National Natural Science Foundation of China (No.81370552, No.81172303), and the Innovation Project of Medical Science and Technology from Nanjing Military Command (No.10MA127)
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