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. 1998 Jul;78(1):62–68. doi: 10.1038/bjc.1998.443

Thymidylate synthase expression and activity: relation to S-phase parameters and 5-fluorouracil sensitivity.

J F Mirjolet 1, M Barberi-Heyob 1, J L Merlin 1, S Marchal 1, M C Etienne 1, G Milano 1, P Bey 1
PMCID: PMC2062951  PMID: 9662252

Abstract

Six human cancer cell lines exhibiting a large range of sensitivity to 5-fluorouracil (5-FU) were evaluated for thymidylate synthase (TS) and p53 gene expression, TS and dihydropyrimidine dehydrogenase (DPD) activity, as well as cell cycle parameters, S-phase fraction (SPF), bromodeoxyuridine labelling index (LI) and S-phase duration (SPD). All these parameters were investigated for 7 days in asynchronously growing cell populations and compared with the cell sensitivity to 5-FU. No significant correlation was found between S-phase parameters and TS gene expression and/or activity. TS activity was higher in proliferating cells; however, it was not significantly higher in rapidly growing cell lines with short SPD. Neither TS gene expression nor activity was found to correlate with 5-FU sensitivity. On the another hand, a statistically significant correlation (P < 0.0001) was observed between LI and SPD and 5-FU sensitivity. The present results suggest that cell cycle parameters such as SPD and/or LI could be better parameters for 5-FU sensitivity prediction than TS gene expression and/or activity. This could be especially informative in cases of concomitant radio-chemotherapy as S-phase parameters are already proposed for hyperfractionated radiotherapy planning.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Aguilar-Santelises M., Rottenberg M. E., Lewin N., Mellstedt H., Jondal M. Bcl-2, Bax and p53 expression in B-CLL in relation to in vitro survival and clinical progression. Int J Cancer. 1996 Apr 22;69(2):114–119. doi: 10.1002/(SICI)1097-0215(19960422)69:2<114::AID-IJC8>3.0.CO;2-3. [DOI] [PubMed] [Google Scholar]
  2. Barberi-Heyob M., Griffon G., Merlin J. L., Weber B. Sequence-dependent growth-inhibitory effects of the in vitro combination of fluorouracil, cisplatin, and dipyridamole. Cancer Chemother Pharmacol. 1993;33(2):163–170. doi: 10.1007/BF00685336. [DOI] [PubMed] [Google Scholar]
  3. Barberi-Heyob M., Weber B., Merlin J. L., Dittrich C., de Bruijn E. A., Luporsi E., Guillemin F. Evaluation of plasma 5-fluorouracil nucleoside levels in patients with metastatic breast cancer: relationships with toxicities. Cancer Chemother Pharmacol. 1995;37(1-2):110–116. doi: 10.1007/BF00685637. [DOI] [PubMed] [Google Scholar]
  4. Beck A., Etienne M. C., Chéradame S., Fischel J. L., Formento P., Renée N., Milano G. A role for dihydropyrimidine dehydrogenase and thymidylate synthase in tumour sensitivity to fluorouracil. Eur J Cancer. 1994;30A(10):1517–1522. doi: 10.1016/0959-8049(94)00216-r. [DOI] [PubMed] [Google Scholar]
  5. Cadman E., Heimer R. Levels of thymidylate synthetase during normal culture growth of L1210 cells. Cancer Res. 1986 Mar;46(3):1195–1198. [PubMed] [Google Scholar]
  6. Chu E., Allegra C. J. The role of thymidylate synthase in cellular regulation. Adv Enzyme Regul. 1996;36:143–163. doi: 10.1016/0065-2571(95)00004-6. [DOI] [PubMed] [Google Scholar]
  7. Chu E., Drake J. C., Koeller D. M., Zinn S., Jamis-Dow C. A., Yeh G. C., Allegra C. J. Induction of thymidylate synthase associated with multidrug resistance in human breast and colon cancer cell lines. Mol Pharmacol. 1991 Feb;39(2):136–143. [PubMed] [Google Scholar]
  8. Chu E., Koeller D. M., Casey J. L., Drake J. C., Chabner B. A., Elwood P. C., Zinn S., Allegra C. J. Autoregulation of human thymidylate synthase messenger RNA translation by thymidylate synthase. Proc Natl Acad Sci U S A. 1991 Oct 15;88(20):8977–8981. doi: 10.1073/pnas.88.20.8977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chu E., Lai G. M., Zinn S., Allegra C. J. Resistance of a human ovarian cancer line to 5-fluorouracil associated with decreased levels of 5-fluorouracil in DNA. Mol Pharmacol. 1990 Sep;38(3):410–417. [PubMed] [Google Scholar]
  10. Conrad A. H. Thymidylate synthetase activity in cultured mammalian cells. J Biol Chem. 1971 Mar 10;246(5):1318–1323. [PubMed] [Google Scholar]
  11. Evans R. M., Laskin J. D., Hakala M. T. Assessment of growth-limiting events caused by 5-fluorouracil in mouse cells and in human cells. Cancer Res. 1980 Nov;40(11):4113–4122. [PubMed] [Google Scholar]
  12. Findlay M. P., Cunningham D., Morgan G., Clinton S., Hardcastle A., Aherne G. W. Lack of correlation between thymidylate synthase levels in primary colorectal tumours and subsequent response to chemotherapy. Br J Cancer. 1997;75(6):903–909. doi: 10.1038/bjc.1997.159. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Güssow D., Rein R., Ginjaar I., Hochstenbach F., Seemann G., Kottman A., Ploegh H. L. The human beta 2-microglobulin gene. Primary structure and definition of the transcriptional unit. J Immunol. 1987 Nov 1;139(9):3132–3138. [PubMed] [Google Scholar]
  14. Harris B. E., Song R., Soong S. J., Diasio R. B. Relationship between dihydropyrimidine dehydrogenase activity and plasma 5-fluorouracil levels with evidence for circadian variation of enzyme activity and plasma drug levels in cancer patients receiving 5-fluorouracil by protracted continuous infusion. Cancer Res. 1990 Jan 1;50(1):197–201. [PubMed] [Google Scholar]
  15. Imam A. M., Crossley P. H., Jackman A. L., Little P. F. Analysis of thymidylate synthase gene amplification and of mRNA levels in the cell cycle. J Biol Chem. 1987 May 25;262(15):7368–7373. [PubMed] [Google Scholar]
  16. Jenh C. H., Rao L. G., Johnson L. F. Regulation of thymidylate synthase enzyme synthesis in 5-fluorodeoxyuridine-resistant mouse fibroblasts during the transition from the resting to growing state. J Cell Physiol. 1985 Jan;122(1):149–154. doi: 10.1002/jcp.1041220122. [DOI] [PubMed] [Google Scholar]
  17. Kaneda S., Takeishi K., Ayusawa D., Shimizu K., Seno T., Altman S. Role in translation of a triple tandemly repeated sequence in the 5'-untranslated region of human thymidylate synthase mRNA. Nucleic Acids Res. 1987 Feb 11;15(3):1259–1270. doi: 10.1093/nar/15.3.1259. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Matherly L. H., Schuetz J. D., Westin E., Goldman I. D. A method for the synchronization of cultured cells with aphidicolin: application to the large-scale synchronization of L1210 cells and the study of the cell cycle regulation of thymidylate synthase and dihydrofolate reductase. Anal Biochem. 1989 Nov 1;182(2):338–345. doi: 10.1016/0003-2697(89)90605-2. [DOI] [PubMed] [Google Scholar]
  19. Pestalozzi B. C., McGinn C. J., Kinsella T. J., Drake J. C., Glennon M. C., Allegra C. J., Johnston P. G. Increased thymidylate synthase protein levels are principally associated with proliferation but not cell cycle phase in asynchronous human cancer cells. Br J Cancer. 1995 Jun;71(6):1151–1157. doi: 10.1038/bjc.1995.225. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Plucinski T. M., Fager R. S., Reddy G. P. Allosteric interaction of components of the replitase complex is responsible for enzyme cross-inhibition. Mol Pharmacol. 1990 Jul;38(1):114–120. [PubMed] [Google Scholar]
  21. Schimke R. T. Gene amplification, drug resistance, and cancer. Cancer Res. 1984 May;44(5):1735–1742. [PubMed] [Google Scholar]
  22. Smith P. K., Krohn R. I., Hermanson G. T., Mallia A. K., Gartner F. H., Provenzano M. D., Fujimoto E. K., Goeke N. M., Olson B. J., Klenk D. C. Measurement of protein using bicinchoninic acid. Anal Biochem. 1985 Oct;150(1):76–85. doi: 10.1016/0003-2697(85)90442-7. [DOI] [PubMed] [Google Scholar]
  23. Storms R. K., Ord R. W., Greenwood M. T., Mirdamadi B., Chu F. K., Belfort M. Cell cycle-dependent expression of thymidylate synthase in Saccharomyces cerevisiae. Mol Cell Biol. 1984 Dec;4(12):2858–2864. doi: 10.1128/mcb.4.12.2858. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Takeishi K., Kaneda S., Ayusawa D., Shimizu K., Gotoh O., Seno T. Nucleotide sequence of a functional cDNA for human thymidylate synthase. Nucleic Acids Res. 1985 Mar 25;13(6):2035–2043. doi: 10.1093/nar/13.6.2035. [DOI] [PMC free article] [PubMed] [Google Scholar]

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