Tissue inhibitor of metalloproteinase-2 inhibits ameloblastoma growth in a new mouse xenograft disease model

J Oral Pathol Med. 2010 Jan;39(1):94-102. doi: 10.1111/j.1600-0714.2009.00812.x. Epub 2009 Nov 6.

Abstract

Background: Ameloblastomas are odontogenic neoplasms characterized by local invasiveness. This study was conducted to develop a new animal model of ameloblastoma and to address the role of tissue inhibitor of metalloproteinase-2 (TIMP-2) and matrix metalloproteinase-2 (MMP-2) in the growth and invasiveness of ameloblastomas.

Method: Donated fresh human ameloblastoma tissue was finely minced, screened, and subcutaneously implanted in three locations on each of 10 BALB/c-nu/nu nude mice. Newly established tumors on each mouse were injected with: (i) transfection reagent; (ii) liposome and transfection reagent; or (iii) liposome, transfection reagent, and the expression plasmid pcDNA3.1(+)/green fluorescent protein (GFP)-TIMP-2. Tumors were monitored for 5 weeks and excised for histopathology, RNA, and protein analyses.

Results: The ameloblastoma xenografts were established with high frequency and contained a variety of typical features, validating this new model system. Xenografts injected with the TIMP-2 expression plasmid showed reduced growth, increased TIMP-2 mRNA and protein, and decreased MMP-2 protein compared with the control groups.

Conclusions: We successfully established a new experimental model of ameloblastoma consisting of subcutaneous human xenografts in nude mice. In addition, we demonstrated the successful introduction of the TIMP-2 gene in tumor xenograft cells in vivo, resulting in xenograft growth inhibition. This growth inhibition may have resulted from TIMP-2 overexpression specifically inhibiting MMP-2 protein expression and activity.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adult
  • Ameloblastoma / drug therapy*
  • Animals
  • Blotting, Western
  • Disease Models, Animal
  • Female
  • Genetic Vectors / therapeutic use
  • Humans
  • Matrix Metalloproteinase 2
  • Matrix Metalloproteinase Inhibitors
  • Mice
  • Mice, Inbred BALB C
  • Mice, Nude
  • Neoplasm Invasiveness
  • Neoplasm Transplantation*
  • Plasmids / therapeutic use
  • Reverse Transcriptase Polymerase Chain Reaction
  • Soft Tissue Neoplasms / drug therapy*
  • Subcutaneous Tissue / surgery
  • Tissue Inhibitor of Metalloproteinase-2 / genetics
  • Tissue Inhibitor of Metalloproteinase-2 / therapeutic use*
  • Transfection
  • Transplantation, Heterologous*
  • Tumor Cells, Cultured

Substances

  • Matrix Metalloproteinase Inhibitors
  • Tissue Inhibitor of Metalloproteinase-2
  • Matrix Metalloproteinase 2
  • Mmp2 protein, mouse