[Effects of nolatrexed on thymidylate synthase protein expression].
OBJECTIVE
To study the dynamic changes in thymidylate synthase (TS) expression in different cell lines treated with nolatrexed.


METHODS
For studying the time course of the action of the drug, LoVo and Lo2 cells were treated separately with nolatrexed at the concentrations of 10 and 75 micromol/L (approximately their respective IC(90) value) for 0 to 60 h, while in the dose response study, treatment with nolatrexed for 36 h was performed with the dose range of 0 to 40 micromol/L for LoVo cells and 0 to 125 micromol/L for Lo2 cell. TS expression was detected by flow cytometry and Western blotting.


RESULTS
The dynamic changes in TS expression with respect of the time course and dose response differed significantly in the two cell lines after nolatrexed treatment. In the time course study, TS expression increased continuously in LoVo cells, as compared with that in the control cells, whereas the TS expression initially decreased and then began to increase gradually 24 h after the treatment, remaining still below the control level after 60 h. In the dose response study, TS expression in LoVo cells gave rise to a peak in the curve, with the peak level approaching the IC(90) value. In Lo2 cells, when the concentration of nolatrexed was blow the IC(90) value, the decrement in TS expression were similar, but as the concentration exceeded the IC(90) value, TS expression decreased dose-dependently.


CONCLUSIONS
Nolatrexed may elicit TS induction in the cells, and the difference in the dynamic changes of TS expression between different cell lines after nolatrexed treatment contributes to different profiles of drug resistance.
Institute of Pharmaceutical Sciences, First Military Medical University, Guangzhou 510515, China.
Yi-lei
YL
Li
Shu-guang
SG
Wu
D002460:Cell Line; D015179:Colorectal Neoplasms / Q000201:enzymology* / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D019008:Drug Resistance, Neoplasm; D022781:Hepatocytes / Q000145:classification / Q000201:enzymology; D006801:Humans; D011799:Quinazolines / Q000494:pharmacology*; D013940:Thymidylate Synthase / Q000037:antagonists & inhibitors / Q000096:biosynthesis* / Q000235:genetics; D014407:Tumor Cells, Cultured
D011799:Quinazolines; D013940:Thymidylate Synthase; C099178:nolatrexed