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

J-F Mirjolet,G Milano,P Bey,J-L Merlin,S Marchal,M Barberi-Heyob,M-C Etienne

doi:10.1038/bjc.1998.443

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.

Highlights

S-phase parameters and thymidylate synthase (TS) gene expression and/or actvity
as we previously demonstrated that cell lines with a shorter doubling time exhibited significantly higher sensitivity to 5-FU (Mirjolet et al 1997). cell cycle kinetic parameters were evaluated using flow cytometry
Results were expressed as relative absorbance to untreated controls. 5-FU concentrations yielding 50% growth inhibition (ICy) were calculated using medium-effect algorithm (Chou and Talalay, 1987) and expressed as mean values of three independent experiments

Summary

Introduction

S-phase parameters and TS gene expression and/or actvity. Neither TS gene expression nor actvity 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 sensifivity predicon 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

Objectives

Findings

Discussion

Conclusion