Antitumor effects of 5-fluorouracil on human colon cancer cell lines: antagonism by levamisole.
BACKGROUND
The addition of levamisole (Lev) to 5-fluorouracil (5-FU) for the adjuvant treatment of stage III colon cancer has been shown to improve 5-year survival in patients. The mechanism of action of Lev remains unknown. Because we showed little in vitro immunological effect of Lev, we asked whether Lev, alone or in combination with 5-FU, had antitumor activity in vitro.


METHODS
Proliferation of COLO-205 and HT-29 colon cancer cells incubated for 2 to 3 days in Lev and 5-FU was measured in 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium colorimetric assays. Cell cycle analysis was performed by treating tumor cells for 6, 24, and 48 h with Lev and 5-FU, staining cells with propidium iodide, and measuring DNA content by flow cytometry.


RESULTS
The addition of Lev to 5-FU did not reduce proliferation below that of 5-FU alone. The inhibitory concentration 50% (IC(50)) for 5-FU was 3.2 x 10(-6) M for COLO-205 and 1.3 x 10(-5) M for HT-29. An IC(50) was not reached for Lev, even at millimolar doses. DNA analysis of cells treated for 48 h revealed significant S-phase accumulation of both HT-29 (from 17% in control cells to 36% in treated cells) and COLO-205 (from 35% in control cells to 59% in treated cells) cell lines at micromolar 5-FU concentrations. In contrast, Lev alone did not affect cell cycle distribution for either cell line. The addition of Lev to 5-FU not only did not augment, but inhibited, the effects of 5-FU.


CONCLUSIONS
Levamisole has no direct cytotoxic effect and no additive or synergistic cytotoxic effect when combined with 5-FU on two colon cancer cell lines. Either the observed clinical effects of Lev treatment occur through an as yet unknown mechanism, require longer treatment periods in vitro to become evident, or the results of clinical trials showing its effectiveness should be carefully reexamined.
Wiebke
Eric A
EA
Department of Surgery, Indiana University School of Medicine and Surgical Service, Roudebush VA Medical Center, Indianapolis, Indiana 46202, USA. ewiebke@iupui.edu
Grieshop
Neil A
NA
Loehrer
Patrick J
PJ
Eckert
George J
GJ
Sidner
Richard A
RA
D000964:Antimetabolites, Antineoplastic / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D002455:Cell Division / Q000187:drug effects; D003110:Colonic Neoplasms / Q000737:chemistry / Q000473:pathology*; D004247:DNA / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D005434:Flow Cytometry; D005472:Fluorouracil / Q000008:administration & dosage / Q000494:pharmacology*; D006801:Humans; D007978:Levamisole / Q000008:administration & dosage / Q000494:pharmacology*; D016196:S Phase; D014407:Tumor Cells, Cultured
D000964:Antimetabolites, Antineoplastic; D007978:Levamisole; D004247:DNA; D005472:Fluorouracil