title: Regulation gene expression of miR200c and ZEB1 positively enhances effect of tumor vaccine B16F10/GPI-IL-21 on inhibition of melanoma growth and metastasis.
abstract: BACKGROUND
  Genetically modified cells have been shown to be one of the most effective tumor vaccine strategies. However, in many cases, such as in melanoma, induction of a potent immune responses against the disease still remains a major challenge. Thus, novel strategies to reinforce tumor vaccine efficacy are needed. Using microRNA (miR) and Zinc-finger E-box binding homeobox (ZEB) have received much attention for potentially regulating tumor progression. To elicit a potent antitumor efficacy against melanoma, we used tumor vaccine in combination with miR200c overexpression or ZEB1 knockdown to assess the efficacy of treatment of murine melanoma.
  
  
  METHODS
  B16F10 cell vaccine expressing interleukin 21 (IL-21) in the glycosylpho- sphatidylinositol (GPI)-anchored form (B16F10/GPI-IL-21) were developed. The vaccine was immunized into mice challenged by B16F10 cells or B16F10 cells stably transduced with lentiviral-miR200c (B16F10/miR200c) or transfected with the ZEB1-shRNA recombinant (B16F10/shZEB1) or the B16F10/GPI-IL-21 vaccine. The immune responses, tumorigenicity and lung metastasis in mice were evaluated, respectively.
  
  
  RESULTS
  The vaccination with B16F10/GPI-IL-21 markedly increased the serum cytokine levels of IFN-γ, TNF-α, IL-4 and decreased TGF-β level as well as augmented the cytotoxicity of splenocytes in immunized mice compared with control mice. In addition, the tumor vaccine B16F10/GPI-IL-21 significantly inhibited the tumor growth and reduced counts of lung metastases in mice challenged by B16F10/GPI-IL-21, B16F10/shZEB1 and B16F10/miR200c respectively compared with the control mice challenged by B16F10 cells. The efficacy mechanisms may involve in reinforcing immune responses, increasing expression of miR200c, E-cadherin and SMAD-7 and decreasing expression of TGF-β, ZEB1, Vimentin and N-cadherin in tumor tissues from the immunized mice.
  
  
  CONCLUSIONS
  These results indicate that the tumor vaccine B16F10/GPI-IL-21 in combination with miR200c overexpression or ZEB1 knockdown effectively inhibited melanoma growth and metastasis a murine model. Such a strategy may, therefore, be used for the clinical trials.
authors.forename: Xiaoying
authors.initials: X
authors.lastname: Wang
authors.forename: Xiangfeng
authors.initials: X
authors.lastname: He
authors.forename: Fengshu
authors.initials: F
authors.lastname: Zhao
authors.forename: Jing
authors.initials: J
authors.lastname: Wang
authors.forename: Hongyi
authors.initials: H
authors.lastname: Zhang
authors.forename: Fangfang
authors.initials: F
authors.lastname: Shi
authors.forename: Yunxia
authors.initials: Y
authors.lastname: Zhang
authors.forename: Kai
authors.initials: K
authors.lastname: Cai
authors.affiliation: Department of Pathogenic Biology and Immunology of Medical School, Southeast University, Nanjing 210009, China. njdoujun@seu.edu.cn.
authors.forename: Jun
authors.initials: J
authors.lastname: Dou
mesh_terms: D000818:Animals; D019496:Cancer Vaccines / Q000276:immunology*; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D016207:Cytokines / Q000097:blood; D003602:Cytotoxicity, Immunologic / Q000235:genetics; D058750:Epithelial-Mesenchymal Transition / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D055785:Gene Knockdown Techniques; D017261:Glycosylphosphatidylinositols / Q000276:immunology*; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D007114:Immunization; D007378:Interleukins / Q000276:immunology*; D051741:Kruppel-Like Transcription Factors / Q000235:genetics* / Q000378:metabolism; D008546:Melanoma, Experimental / Q000097:blood / Q000235:genetics / Q000276:immunology* / Q000473:pathology*; D051379:Mice; D008810:Mice, Inbred C57BL; D035683:MicroRNAs / Q000235:genetics* / Q000378:metabolism; D009362:Neoplasm Metastasis / Q000235:genetics / Q000276:immunology; D012878:Skin Neoplasms / Q000097:blood / Q000235:genetics / Q000276:immunology / Q000473:pathology; D013154:Spleen / Q000378:metabolism / Q000473:pathology; D014161:Transduction, Genetic; D014162:Transfection; D000071799:Zinc Finger E-box-Binding Homeobox 1; D000098546:Interleukin-21
chemical_list: D019496:Cancer Vaccines; D016207:Cytokines; D017261:Glycosylphosphatidylinositols; D018398:Homeodomain Proteins; D007378:Interleukins; D051741:Kruppel-Like Transcription Factors; D035683:MicroRNAs; C543719:Mirn200 microRNA, mouse; C519422:ZEB1 protein, mouse; D000071799:Zinc Finger E-box-Binding Homeobox 1; D000098546:Interleukin-21