Enhanced cytotoxicity with interleukin-1α and 5-fluorouracil in HCT116 colon cancer cells

F. J. Geoffroy, C. J. Allegra, B. Sinha, Jean L Grem

Research output: Contribution to journalArticle

36 Citations (Scopus)

Abstract

Recombinant human interleukin-1α (rIL-1α), at concentrations that were not growth-inhibitory when given alone (100-10,000 U/ml), enhanced the growth inhibition resulting from a 72-h fluorouracil (FUra) exposure in HCT116 colon cancer cells. Median-effect analysis of clonogenic assays indicated that rIL- 1α, given 24 h prior to and following a 24-h exposure to FUra, increased lethality in a more than additive fashion. rIL-1α did not appear to significantly affect [3H]-FUra metabolism, total [3H]-FUra-RNA incorporation or RNA retention after drug removal, inhibition of thymidylate synthase, or thymidine triphosphate pool depletion. During continuous exposure to rIL-1α, transient stimulation of RNA and DNA synthesis was observed at 72 h, with a return to normal by 96 h. A 24-h exposure to 10 μM FUra altered the elution profile of newly synthesized DNA as monitored by pH step alkaline elution. An accumulation of lower-MW single-stranded DNA species was noted with FUra compared to control, accompanied by a significantly decreased proportion of DNA retained on the polycarbonate filter: 10% retained vs. 32% for control (P = 0.01). A 48-h exposure to rIL- 1α alone did not affect the elution profile of nascent DNA species, nor did it enhance the effects of FUra. Although FUra did not appreciably affect pulse [3H]-uridine incorporation into RNA for the initial 8-24 h of FUra exposure, progressive inhibition of net RNA synthesis was observed thereafter. FUra prevented the stimulatory effect of rIL-1α on RNA synthesis, and net RNA synthesis was significantly inhibited (by 64-79% after 72 and 96 h) with the combination compared to rIL-1α alone. Continuous exposure to 10 μM thymidine did not rescue cells from the lethality of FUra alone or the combination of FUra plus rIL-1α suggesting that depletion of deoxythymidine triphosphate as a consequence of thymidylate synthase inhibition was not the most important component of FUra toxicity. In contrast, 1 mM uridine provided partial protection against the toxicity of FUra alone or with rIL-1α. Although uridine did not affect FUra metabolism, it decreased FUra-RNA incorporation by 42-60%, presumably as a consequence of the 2-fold expansion of UTP pools. [125I]-rIL-1α binding was nonspecific; with a 24-h exposure, however, internalized [125I]-rIL-1α exceeded cell surface-bound material by 2-fold. Concurrent exposure to 10 μM FUra increased the binding and internalization of [125I]-rIL-1α at 37°C by 2.7-fold. While the precise mechanism of interaction between FUra and IL-1α is unclear, the data suggest that RNA-directed cytotoxicity by FUra is contributory.

Original languageEnglish (US)
Pages (from-to)581-591
Number of pages11
JournalOncology Research
Volume6
Issue number12
StatePublished - Dec 1 1994

Fingerprint

Interleukin-5
Interleukin-1
Fluorouracil
Colonic Neoplasms
RNA
Uridine
Thymidylate Synthase
polycarbonate
DNA
Uridine Triphosphate
Single-Stranded DNA

Keywords

  • colorectal carcinoma
  • cytokine-drug interaction
  • fluorouracil
  • interleukin-1α

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

Enhanced cytotoxicity with interleukin-1α and 5-fluorouracil in HCT116 colon cancer cells. / Geoffroy, F. J.; Allegra, C. J.; Sinha, B.; Grem, Jean L.

In: Oncology Research, Vol. 6, No. 12, 01.12.1994, p. 581-591.

Research output: Contribution to journalArticle

Geoffroy, F. J. ; Allegra, C. J. ; Sinha, B. ; Grem, Jean L. / Enhanced cytotoxicity with interleukin-1α and 5-fluorouracil in HCT116 colon cancer cells. In: Oncology Research. 1994 ; Vol. 6, No. 12. pp. 581-591.
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AU - Grem, Jean L

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N2 - Recombinant human interleukin-1α (rIL-1α), at concentrations that were not growth-inhibitory when given alone (100-10,000 U/ml), enhanced the growth inhibition resulting from a 72-h fluorouracil (FUra) exposure in HCT116 colon cancer cells. Median-effect analysis of clonogenic assays indicated that rIL- 1α, given 24 h prior to and following a 24-h exposure to FUra, increased lethality in a more than additive fashion. rIL-1α did not appear to significantly affect [3H]-FUra metabolism, total [3H]-FUra-RNA incorporation or RNA retention after drug removal, inhibition of thymidylate synthase, or thymidine triphosphate pool depletion. During continuous exposure to rIL-1α, transient stimulation of RNA and DNA synthesis was observed at 72 h, with a return to normal by 96 h. A 24-h exposure to 10 μM FUra altered the elution profile of newly synthesized DNA as monitored by pH step alkaline elution. An accumulation of lower-MW single-stranded DNA species was noted with FUra compared to control, accompanied by a significantly decreased proportion of DNA retained on the polycarbonate filter: 10% retained vs. 32% for control (P = 0.01). A 48-h exposure to rIL- 1α alone did not affect the elution profile of nascent DNA species, nor did it enhance the effects of FUra. Although FUra did not appreciably affect pulse [3H]-uridine incorporation into RNA for the initial 8-24 h of FUra exposure, progressive inhibition of net RNA synthesis was observed thereafter. FUra prevented the stimulatory effect of rIL-1α on RNA synthesis, and net RNA synthesis was significantly inhibited (by 64-79% after 72 and 96 h) with the combination compared to rIL-1α alone. Continuous exposure to 10 μM thymidine did not rescue cells from the lethality of FUra alone or the combination of FUra plus rIL-1α suggesting that depletion of deoxythymidine triphosphate as a consequence of thymidylate synthase inhibition was not the most important component of FUra toxicity. In contrast, 1 mM uridine provided partial protection against the toxicity of FUra alone or with rIL-1α. Although uridine did not affect FUra metabolism, it decreased FUra-RNA incorporation by 42-60%, presumably as a consequence of the 2-fold expansion of UTP pools. [125I]-rIL-1α binding was nonspecific; with a 24-h exposure, however, internalized [125I]-rIL-1α exceeded cell surface-bound material by 2-fold. Concurrent exposure to 10 μM FUra increased the binding and internalization of [125I]-rIL-1α at 37°C by 2.7-fold. While the precise mechanism of interaction between FUra and IL-1α is unclear, the data suggest that RNA-directed cytotoxicity by FUra is contributory.

AB - Recombinant human interleukin-1α (rIL-1α), at concentrations that were not growth-inhibitory when given alone (100-10,000 U/ml), enhanced the growth inhibition resulting from a 72-h fluorouracil (FUra) exposure in HCT116 colon cancer cells. Median-effect analysis of clonogenic assays indicated that rIL- 1α, given 24 h prior to and following a 24-h exposure to FUra, increased lethality in a more than additive fashion. rIL-1α did not appear to significantly affect [3H]-FUra metabolism, total [3H]-FUra-RNA incorporation or RNA retention after drug removal, inhibition of thymidylate synthase, or thymidine triphosphate pool depletion. During continuous exposure to rIL-1α, transient stimulation of RNA and DNA synthesis was observed at 72 h, with a return to normal by 96 h. A 24-h exposure to 10 μM FUra altered the elution profile of newly synthesized DNA as monitored by pH step alkaline elution. An accumulation of lower-MW single-stranded DNA species was noted with FUra compared to control, accompanied by a significantly decreased proportion of DNA retained on the polycarbonate filter: 10% retained vs. 32% for control (P = 0.01). A 48-h exposure to rIL- 1α alone did not affect the elution profile of nascent DNA species, nor did it enhance the effects of FUra. Although FUra did not appreciably affect pulse [3H]-uridine incorporation into RNA for the initial 8-24 h of FUra exposure, progressive inhibition of net RNA synthesis was observed thereafter. FUra prevented the stimulatory effect of rIL-1α on RNA synthesis, and net RNA synthesis was significantly inhibited (by 64-79% after 72 and 96 h) with the combination compared to rIL-1α alone. Continuous exposure to 10 μM thymidine did not rescue cells from the lethality of FUra alone or the combination of FUra plus rIL-1α suggesting that depletion of deoxythymidine triphosphate as a consequence of thymidylate synthase inhibition was not the most important component of FUra toxicity. In contrast, 1 mM uridine provided partial protection against the toxicity of FUra alone or with rIL-1α. Although uridine did not affect FUra metabolism, it decreased FUra-RNA incorporation by 42-60%, presumably as a consequence of the 2-fold expansion of UTP pools. [125I]-rIL-1α binding was nonspecific; with a 24-h exposure, however, internalized [125I]-rIL-1α exceeded cell surface-bound material by 2-fold. Concurrent exposure to 10 μM FUra increased the binding and internalization of [125I]-rIL-1α at 37°C by 2.7-fold. While the precise mechanism of interaction between FUra and IL-1α is unclear, the data suggest that RNA-directed cytotoxicity by FUra is contributory.

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