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Japanese Journal of Cancer Research : Gann logoLink to Japanese Journal of Cancer Research : Gann
. 1999 Apr;90(4):454–459. doi: 10.1111/j.1349-7006.1999.tb00769.x

Enhancement of the Anti‐tumor Effect of 5′‐Deoxy‐5‐fluorouridine by Transfection of Thymidine Phosphorylase Gene into Human Colon Cancer Cells

Hiroki Kanyama 1, Naohiro Tomita 1, Tomoki Yamano 1, Yasuo Miyoshi 1, Masayuki Ohue 1, Yoshiyuki Fujiwara 1, Mitsugu Sekimoto 1, Isao Sakita 1, Yasuhiro Tamaki 1, Morito Monden 1
PMCID: PMC5926090  PMID: 10363585

Abstract

Thymidine phosphorylase (dThdPase) is an enzyme that converts 5′‐deoxy‐5‐fluorouridine (5′DFUR) to the toxic substance 5‐fluorouracil (5‐FU); it is also known to be a platelet‐derived endothelial cell growth factor. In order to investigate the feasibility of suicide gene therapy against colorectal cancer by means of the combination of 5′DFUR and the converting enzyme dThdPase, we transfected the dThdPase gene into the human colon cancer cell line SW480 and analyzed the growth pattern as well as the sensitivity to 5‐FU or 5′DFUR of the dThdPase‐transfected cells. The 50% inhibition (IC50) values of 5‐FU against the SW480 parental cells, control vector‐transfected cells SW480/V1, and dThdPase‐transfected cells SW480/dThdPase were approximately 4.9, 6.3, and 2.9 μM, respectively. The IC50 of SW480/dThdPase was lower than that of SW480 or SW480/V1, although the differences were not statistically significant. The IC50 values of 5′DFUR for SW480, SW480/V1, and SW480/dThdPase were approximately 300, 330, and 3.2 αM, respectively. The sensitivity to 5′DFUR of SW480/dThdPase was increased by about 100‐fold compared with that of SW480 or SW480/V1. With only 10% transfection efficacy, a high enough sensitivity to 5′DFUR was obtained to suppress the cell growth, indicating that a strong bystander effect was induced by this system. The in vivo growth of the s.c. transplanted SW480/dThdPase tumor in nude mice was significantly suppressed by i.p. injection of 5′DFUR compared with that in control mice that received phosphate‐buffered saline (PBS) treatment. These results suggest that gene therapy using the combination of 5′DFUR and the dThdPase gene may be a useful approach for treatment of colon cancer.

Keywords: dThdPase, Converting enzyme, 5′DFUR, Colon cancers, Suicide gene therapy

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E‐mail: cba77840@pop06.odn.ne.jp

REFERENCES

  • 1).Mullen , C. A. , Kilstrup , M. and Blaese , R. M.Transfer of the bacterial gene for cytosine deaminase to mammalian cells confers lethal sensitivity to 5‐fluorocytosine: a negative selection system . Proc. Natl. Acad. Sci. USA , 89 , 33 – 37 ( 1992. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 2).Mullen , C. A. , Coale , M. M. , Lowe , R. and Blaese , R. M.Tumors expressing the cytosine deaminase suicide gene can be eliminated in vivo with 5‐fluorocytosine and induce protective immunity to wild type tumor . Cancer Res. , 54 , 1503 – 1506 ( 1994. ). [PubMed] [Google Scholar]
  • 3).Sugaya , S. , Fujita , K. , Kikuchi , A. , Ueda , H. , Takakuwa , K. , Shoji , K. and Tanaka , K.Inhibition of tumor growth by direct intratumoral gene transfer of herpes simplex virus thymidine kinase gene with DNA‐liposome complex . Hum. Gene Ther. , 7 , 223 – 230 ( 1996. ). [DOI] [PubMed] [Google Scholar]
  • 4).Bodycote , J. and Wolff , S.Metabolic breakdown of [3H]thymidine and the inability to measure human lymphocyte proliferation by incorporation of radioactivity . Proc. Natl. Acad. Sci. USA , 83 , 4749 – 4753 ( 1986. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 5).Birine , G. D. , Kroeger , H. and Heidelberger , C.Studies of fluorinated pyrimidines. XVIII. The degradation of 5‐flu‐oro‐2′‐deoxyuridine and related compounds by nucleoside phosphorylase . Biochemistry , 2 , 566 – 572 ( 1963. ). [DOI] [PubMed] [Google Scholar]
  • 6).Krenitsky , T. A. and Koszalka , J. V.Purine nucleoside synthesis, an efficient method employing nucleoside phosphorylases . Biochemistry , 20 , 3615 – 3621 ( 1981. ). [DOI] [PubMed] [Google Scholar]
  • 7).Zimmerman , M. and Seidenberg , J.Deoxyribosyl transfer. I. Thymidine phosphorylase and nucleoside deoxyribosyl‐transferase in normal and mucosal tissues . J. Biol. Chem. , 239 , 2618 – 2621 ( 1964. ). [PubMed] [Google Scholar]
  • 8).Marsh , J. C. and Perry , S.Thymidine catabolism by normal and leukemic human leucocytes . J. Clin. Invest. , 43 , 267 – 278 ( 1964. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 9).Shaw , T. , Smillie , R. H. , Miller , A. E. and Macphee , D. G.The role of blood platelets in nucleoside metabolism: regulation of platelet thymidine phosphorylase . Mutat. Res. , 200 , 117 – 131 ( 1988. ). [DOI] [PubMed] [Google Scholar]
  • 10).Yoshimura , A. , Kuwazuru , Y. , Furukawa , T. , Yoshida , H. , Yamada , K. and Akiyama , K.Purification and tissue distribution of human thymidine phosphorylase: high expression in lymphocytes, reticulocytes and tumors . Biochim. Biophys. Acta , 1034 , 107 – 113 ( 1990. ). [DOI] [PubMed] [Google Scholar]
  • 11).Luccioni , C. , Beaumatin , J. , Bardot , V. and Lefrancois , D.Pyrimidine nucleotide metabolism in human colon carcinomas; comparison of normal tissues, primary tumors and xenografts . Int. J. Cancer , 58 , 517 – 522 ( 1994. ). [DOI] [PubMed] [Google Scholar]
  • 12).Peters , G. J. , van Groeningen , C. J. , Laurensse , E. J. and Pinedo , H. M.A comparison of 5‐fluorouracil metabolism in human colorectal cancer and colon mucosa . Cancer , 68 , 1903 – 1909 ( 1991. ). [DOI] [PubMed] [Google Scholar]
  • 13).Haba , A. , Monden , T. , Sekimoto , M. , Ikeda , K. , Kanyama , H. and Monden , M.PyNPase expression in human colon cancer . Cancer Lett. , 122 , 85 – 92 ( 1998. ). [DOI] [PubMed] [Google Scholar]
  • 14).Ishikawa , M. , Shiga , M. , Sasamoto , K. and Mizoguchi , M.A new sulfonated tetrazolium salt that produces a highly water‐soluble formazan dye . Chem. Pharm. Bull. , 41 , 1118 – 1122 ( 1993. ). [Google Scholar]
  • 15).Haraguchi , M. , Furukawa , T. , Sumizawa , T. and Akiyama , S.Sensitivity of human KB cells expressing platelet‐derived endothelial cell growth factor to pyrimidine antimetabolites . Cancer Res. , 53 , 5680 – 5682 ( 1993. ). [PubMed] [Google Scholar]
  • 16).Patterson , A. V. , Zhang , H. , Moghaddam , A. , Bicknell , R. , Talbot , D. C. , Stratford , I. J. and Harris , A. L.Increased sensitivity to the prodrug 5′‐deoxy‐5‐fluorouridine and modulation of 5‐fluoro‐2′‐deoxyuridine sensitivity in MCF‐7 cells transfected with thymidine phosphorylase . Br. J. Cancer , 72 , 669 – 675 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 17).Schwartz , E. L. , Baptiste , N. , O'Connor , C. J. , Wadler , S. and Otter , B. A.Potentiation of 5‐fluorouracil in colon carcinoma cells by the combination of interferon and deoxyribonucleotides results from complementary effects on thymidine phosphorylase . Cancer Res. , 54 , 1472 – 1478 ( 1994. ). [PubMed] [Google Scholar]
  • 18).Freeman , S. M. , Abboud , C. N. and Whartenby , K. A.The bystander effect: tumor regression when a fraction of the tumor mass is genetically modified . Cancer Res. , 53 , 1503 – 1506 ( 1994. ). [PubMed] [Google Scholar]
  • 19).Kuriyama , S. , Matui , K. , Sakamoto , T. , Nakatani , T. , Ikenaga , K. and Tsujii , T.Bacterial cytosine deaminase suicide gene transduction renders hepatocellular carcinoma sensitive to the prodrug 5‐fluorocytosine . Int. Hepatol. Commun. , 4 , 72 – 79 ( 1995. ). [Google Scholar]
  • 20).Tomita , N. , Jiang , W. , Hibshoosh , H. , Warburton , D. , Kahn , S. M. and Weinstein , I. B.Isolation and characterization of a highly malignant variant of the SW480 human colon cancer cell line . Cancer Res. , 52 , 6840 – 6847 ( 1992. ). [PubMed] [Google Scholar]
  • 21).Moghaddam , A. , Zhang , H. T. , Fan , T. P. , Hu , D. E. , Lees , V. C. , Turley , H. , Fox , S. B. , Gatter , K. C. , Harris , A. L. and Bicknell , R.Thymidine phosphorylase is angiogenic and promotes tumor growth . Proc. Natl. Acad. Sci. USA , 92 , 998 – 1002 ( 1995. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
  • 22).Furukawa , T. , Yoshimura , A. , Sumizawa , T. , Haraguchi , M. and Akiyama , S. I.Angiogenic factor . Nature , 356 , 668 ( 1992. ). [DOI] [PubMed] [Google Scholar]
  • 23).Miyazono , K. , Okabe , T. , Urabe , A. , Takaku , F. and Heldin , C. H.Purification and properties of an endothelial cell growth factor from human platelets . J. Biol. Chem. , 262 , 4098 – 4103 ( 1987. ). [PubMed] [Google Scholar]
  • 24).Ishikawa , F. , Miyazono , K. , Hellman , U. , Drexler , H. , Wernstedt , C. , Hagiwara , K. , Usuki , K. , Takaku , F. , Risau , W. and Heldin , C.H. Identification of angiogenic activity and the cloning and expression of platelet‐derived endothelial cell growth factor . Nature , 338 , 557 – 562 ( 1989. ). [DOI] [PubMed] [Google Scholar]
  • 25).Takebayashi , Y. , Yamada , K. , Sumizawa , T. , Miyadera , K. , Aikou , T. and Akiyama , S.Clinicopathological significance of an angiogenic enzyme, thymidine phosphorylase (dThdPase), in colorectal carcinoma . Proc. Am. Assoc. Cancer Res. , 36 , 90 ( 1995. ). [Google Scholar]

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