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Establishment of a Tumor-bearing Mouse Model Stably Expressing Human Tumor Antigens Survivin and MUC1 VNTRs

Establishment of a Tumor-bearing Mouse Model Stably Expressing Human Tumor Antigens Survivin and MUC1 VNTRs
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摘要 The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1(MUC1)(VNTRs),namely,VR1012-S and VR1012-VNTR(VNTR=variable number of tandem repeat),were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line(B16) stably expressing survivin and MUC1 VNTRs(MS + B16) was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS + B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs(MS) expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS + B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×l0 5 MS + B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline(PBS),a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs. The eukaryotic vectors VR1012 expressing survivin or 33 tandem repeats of human mucin 1(MUC1)(VNTRs),namely,VR1012-S and VR1012-VNTR(VNTR=variable number of tandem repeat),were constructed by cloning survivin and VNTR genes into VR1012,respectively.The eukaryotic vector pEGFP expressing survivin and MUC1 VNTRs fusion gene pEGFP-MS was also constructed.Mouse melanoma cell line(B16) stably expressing survivin and MUC1 VNTRs(MS + B16) was established by Lipofectamine-mediated transfection of pEGFP-MS into B16 cells.EGFP expression in MS + B16 cells was observed using a fluorescent microscope and survivin and MUC1 VNTRs(MS) expression was confirmed by means of Western blot analysis.A syngenic graft tumor model was generated by subcutaneous injection of MS + B16 cells into C57/BL6 mice and tumor size increased rapidly with time in a cell number dependent manner.After the third immunization,mice were challenged subcutaneously with 5×l0 5 MS + B16 cells.Compared with that of the negative control immunized with phosphate-buffered saline(PBS),a significant reduction of tumor growth was observed in groups immunized with survivin plasmid DNA and MUC1 VNTRs plasmid DNA.Thus,the suppression of subcutaneous tumor was antigen-specific.This model is useful for the development of tumor vaccines targeting survivin and MUCI VNTRs.
出处 《Chemical Research in Chinese Universities》 SCIE CAS CSCD 2012年第2期259-263,共5页 高等学校化学研究(英文版)
基金 Supported by the National Natural Science Foundation of China(No.30872396) the Scientific Research Foundation of Jilin Province,China(Nos.20080709,200905169) the Jilin University Basic Research Project,China(No.200903255)
关键词 SURVIVIN MUC1 variable-number tandem repeat(MUC1 VNTR) Tumor antigen Tumor model Tumor vaccine Survivin MUC1 variable-number tandem repeat(MUC1 VNTR) Tumor antigen Tumor model Tumor vaccine
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