Anti-tumorigenic and Invasive Activity of Colon Cancer Cells Transfected with the Retroviral Vector Encoding Tissue Inhibitor of Metalloproteinase-2

레트로바이러스를 이용한 Tissue Inhibitor of Metalloproteinase-2 유전자 발현이 대장암 세포의 전이 및 종양형성에 미치는 영향

  • 오일웅 (식품의약품안전청 생명공학의약품과) ;
  • 정자영 (식품의약품안전청 생명공학의약품) ;
  • 장석기 (식품의약품안전청 생명공학의약품) ;
  • 이수해 (식품의약품안전청 생명공학의약품) ;
  • 김연수 (배제대학) ;
  • 손여원 (식품의약품안전청 생명공학의약품과)
  • Published : 2004.06.01

Abstract

Matrix metalloproteinases (MMPs) and tissue inhibitors of MMPs (TIMPs) playa key role in tumor invasion and metastasis. As an inhibitor of MMP-2, TIMP-2 is known to block both the invasive and metastatic behavior of cancer cells, and decrease tumor growth activity. We performed this study to investigate the effects of TIMP-2 over-expression induced by retroviral mediated gene transfer in vitro and in vivo. The human colon cancer cell line SW480 was transfected with the retroviral vector encoding TIMP-2. The effects of TIMP-2 over-expression were analyzed by invasion assay and gelatinase activity test in colon cancer cells and tumorigencity in nude mice. In evaluation of the transfection efficiency of the retroviral vector encoding TIMP-2 in colon cancer cells, we confirmed up-regulation of TIMP-2 expression dependent on the time of cell culture. In addition, inhibition of MMP-2 expression in SW480/TIMP-2 was shown by gelatin zymography. In the in vitro invasion assay SW480/TIMP-2 inhibited the invasiveness on matrigel coated with collagen. To determine whether TIMP-2 can modulate in vivo tumorigenicity and metastasis, SW480/TIMP-2 cells were injected subcutaneously in nude mice. The tumor mass formation of SW480/TIMP-2 cells in nude mice was markedly decreased compared to nontransfected cancer cells. These results showed that colon cancer cells transfected with the retroviral vector encoding TIMP-2 inhibits the invasiveness in vitro and tumorigenicity in vivo.

Keywords

References

  1. Sporn, M. B. : The war on cancer. Lancet 347, 1377 (1996) https://doi.org/10.1016/S0140-6736(96)91015-6
  2. Liotta, L. A., Tryggvason,K., Garbisa, S., Hart, I., Foltz, C. M. and Shafdie, S. : Metastatic potential correlates with enzymatic degradation of basement membrane collagen. Nature 284, 67 (1980) https://doi.org/10.1038/284067a0
  3. Matrisian, L. M. : The matrix degrading metalloproteinases. Bioassays 14, 455 (1992) https://doi.org/10.1002/bies.950140705
  4. Li, H., Grenet, C., Opolon, P., Menashi, S., Soria, C. and Yeh, P. et al. : AdTIMP-2 inhibits tumor growth, angiogenesis, and metastasis, and prolongs survival in mice. Hum. Gene Ther. 12, 515 (2001) https://doi.org/10.1089/104303401300042429
  5. Rigg, A. S. and Lemoine, N. R. : Adenoviral delivery of TIMP1 and TIMP2 can modify the invasive behavior of pancreatic cancer and can have a significant antitumor effect in vivo. Cancer Gene Ther. 8, 869 (2001) https://doi.org/10.1038/sj.cgt.7700387
  6. Imren, S., Kohn, D. B., Shimada, H., Blavier, L. and DeClerk, Y. A. : Overexpression of tissue inhibitor of metalloproteinase-2 retroviral-mediated gene transfer in vivo inhibits tumor growth and invasion. Cancer Res. 56, 2891 (1996)
  7. Ahn, S. M., Jeong, S. J., Kim, Y. S., Sohn, Y. W. and Moon, A.: Retroviral delivery of TIMP-2 inhibits H-ras-induced migration and invasion in MCF10A human breast epithelial cells. Cancer Letters 207, 49 (2004)
  8. Pohar, N., Godenschwege, T. A. and Buchner, E. : Invertebrate tissue inhibitor of metalloproteinase structure and nested gene organization within the synapsin locus is conserved from Drosohila to human. Genomics 57, 293 (1999) https://doi.org/10.1006/geno.1999.5776
  9. Declerck, Y. A., Perez, H. S., Boone, T. C., Langley, K. E. and Taylor, S. M. : Inhibition of invasion and metastasis in cells transfected with an inhibitor of metalloproteinases. Cancer Res. 52, 701 (1992)
  10. Ponton, A. B., Coulombe and Skup, D.: Decreased expression of tissue inhibitor of the metalloproteinases in metastatic tumor cells leading to increased levels of collagenase activity. Cancer Res. 51, 2138 (1991)
  11. Yen, N., Ioannides, C. G., Xu, K., Swisher, S. G., Lawrence, D. D., Kemp, B. L., Walsh, G. L. and Roth, J. A. : Cellular and humoral immune responses to adenovirus and p53 protein antigens in patients following intratumoral injection of an adenovirus vector expressing wild-type. Cancer Gene Ther 7, 530 (2000) https://doi.org/10.1038/sj.cgt.7700138
  12. Kim, Y. S. and Risser, R.: TAR-independent transactivation of the murine cytomegalovirus major immediate early promoter by the Tat protein. J. Virol. 67, 239 (1993)
  13. Kim, T., Leibfried-Rutledge, M. L. and First, N. L. : Gene transfer in bovine blastocysts using replication-defective retroviral vectors packaged with gibbon ape leukemia virus envelopes. Mol. Reprod. Develop. 35, 105 (1993) https://doi.org/10.1002/mrd.1080350202
  14. Donahue. R. E. Kessler, S. W., Bonkie, D., Mcdonagh, K., Dunbar, C., Goodman, D., Agricola, B., Byrne, E., Ratteld. M., Moen, R., Bacher,J.,Zsebo, K. M. and Nienhuis, A. W.: Helper virus induction T cell lymphoma in nonhuman primates after retroviral mediated gene transfer. J. Exp. Med. 176, 1125 (1992) https://doi.org/10.1084/jem.176.4.1125
  15. Wilson, C. A. and Miller, A. E.: Evaluation of recom-mendations for replication-competent retrovirus testing associated with use of retroviral vectors. Hum. Gene Ther. 8, 869 (1997) https://doi.org/10.1089/hum.1997.8.7-869
  16. Bauer, T. R., Jr., Miller, A. D. and Hickstein, D. D. : Improved transfer of the leukocyte integrin CD18 subunit into hematopoietic cell lines by using retroviral vectors having a gibbon ape leukemia virus envelope. Blood 86, 2379 (1995)
  17. Bunnell, B. A., Muul, L. M., Donahue, R. E., Blaese, R. M. and Morgan, R. A. : High-efficiency retroviral-mediated gene transfer into human and non-human primate peripheral blood lymphocytes. PNAS 92, 7739 (1995) https://doi.org/10.1073/pnas.92.17.7739
  18. Arii, S., Ishigami, S., Mori, A., Onodera, G. and Imamura, M. : Implication of VEGF and MMPs in hepatic metastasis of human colon cancer. Nippon. Geka. Gakkai. Zasshi. 99, 436 (1998)
  19. Masuda, H. and Aoki, H. : Host expression of matrix metalloproteinase-2 and tissue inhibitor of metalloproteinase2 in normal colon tissue affects metastatic potential of colorectal cancer. Dis. Colon Rectum 42, 393 (1999)