Antitumor effects of 5-fluorouracil on human colon cancer cell lines: Antagonism by levamisole

Eric A. Wiebke, Neil A. Grieshop, Patrick Loehrer, George J. Eckert, Richard A. Sidner

Research output: Contribution to journalArticle

15 Citations (Scopus)

Abstract

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% (IC50) for 5-FU was 3.2 × 10-6 M for COLO-205 and 1.3 × 10-5 M for HT-29. An IC50 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.

Original languageEnglish
Pages (from-to)63-69
Number of pages7
JournalJournal of Surgical Research
Volume111
Issue number1
DOIs
StatePublished - May 1 2003

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Levamisole
Fluorouracil
Colonic Neoplasms
Cell Line
Inhibitory Concentration 50
Cell Cycle
Propidium
DNA
S Phase
Serotonin
Flow Cytometry

Keywords

  • 5-fluorouracil
  • Cell lines
  • Colon cancer
  • Levamisole

ASJC Scopus subject areas

  • Surgery

Cite this

Antitumor effects of 5-fluorouracil on human colon cancer cell lines : Antagonism by levamisole. / Wiebke, Eric A.; Grieshop, Neil A.; Loehrer, Patrick; Eckert, George J.; Sidner, Richard A.

In: Journal of Surgical Research, Vol. 111, No. 1, 01.05.2003, p. 63-69.

Research output: Contribution to journalArticle

Wiebke, Eric A. ; Grieshop, Neil A. ; Loehrer, Patrick ; Eckert, George J. ; Sidner, Richard A. / Antitumor effects of 5-fluorouracil on human colon cancer cell lines : Antagonism by levamisole. In: Journal of Surgical Research. 2003 ; Vol. 111, No. 1. pp. 63-69.
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title = "Antitumor effects of 5-fluorouracil on human colon cancer cell lines: Antagonism by levamisole",
abstract = "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{\%} (IC50) for 5-FU was 3.2 × 10-6 M for COLO-205 and 1.3 × 10-5 M for HT-29. An IC50 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.",
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T1 - Antitumor effects of 5-fluorouracil on human colon cancer cell lines

T2 - Antagonism by levamisole

AU - Wiebke, Eric A.

AU - Grieshop, Neil A.

AU - Loehrer, Patrick

AU - Eckert, George J.

AU - Sidner, Richard A.

PY - 2003/5/1

Y1 - 2003/5/1

N2 - 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% (IC50) for 5-FU was 3.2 × 10-6 M for COLO-205 and 1.3 × 10-5 M for HT-29. An IC50 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.

AB - 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% (IC50) for 5-FU was 3.2 × 10-6 M for COLO-205 and 1.3 × 10-5 M for HT-29. An IC50 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.

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