A systems biology approach identifies SART1 as a novel determinant of both 5-fluorouracil and SN38 drug resistance in colorectal cancer

Wendy L. Allen, Leanne Stevenson, Vicky M. Coyle, Puthen V. Jithesh, Irina Proutski, Gail Carson, Michael A. Gordon, Heinz Josef D Lenz, Sandra Van Schaeybroeck, Daniel B. Longley, Patrick G. Johnston

Research output: Contribution to journalArticle

19 Citations (Scopus)

Abstract

Chemotherapy response rates for advanced colorectal cancer remain disappointingly low, primarily because of drug resistance, so there is an urgent need to improve current treatment strategies. To identify novel determinants of resistance to the clinically relevant drugs 5-fluorouracil (5-FU) and SN38 (the active metabolite of irinotecan), transcriptional profiling experiments were carried out on pretreatment metastatic colorectal cancer biopsies and HCT116 parental and chemotherapy-resistant cell line models using a disease-specific DNA microarray. To enrich for potential chemoresistance-determining genes, an unsupervised bioinformatics approach was used, and 50 genes were selected and then functionally assessed using custom-designed short interfering RNA (siRNA) screens. In the primary siRNA screen, silencing of 21 genes sensitized HCT116 cells to either 5-FU or SN38 treatment. Three genes (RAPGEF2, PTRF, and SART1) were selected for further analysis in a panel of 5 colorectal cancer cell lines. Silencing SART1 sensitized all 5 cell lines to 5-FU treatment and 4/5 cell lines to SN38 treatment. However, silencing of RAPGEF2 or PTRF had no significant effect on 5-FU or SN38 sensitivity in the wider cell line panel. Further functional analysis of SART1 showed that its silencing induced apoptosis that was caspase-8 dependent. Furthermore, silencing of SART1 led to a downregulation of the caspase-8 inhibitor, c-FLIP, which we have previously shown is a key determinant of drug resistance in colorectal cancer. This study shows the power of systems biology approaches for identifying novel genes that regulate drug resistance and identifies SART1 as a previously unidentified regulator of c-FLIP and drug-induced activation of caspase-8. Mol Cancer Ther; 11(1); 119-31.

Original languageEnglish
Pages (from-to)119-131
Number of pages13
JournalMolecular Cancer Therapeutics
Volume11
Issue number1
DOIs
Publication statusPublished - Jan 2012
Externally publishedYes

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Systems Biology
Drug Resistance
Fluorouracil
Colorectal Neoplasms
Caspase 8
Cell Line
irinotecan
Small Interfering RNA
Genes
HCT116 Cells
Drug Therapy
Caspase Inhibitors
Gene Silencing
Therapeutics
Oligonucleotide Array Sequence Analysis
Computational Biology
Down-Regulation
Apoptosis
Biopsy
Pharmaceutical Preparations

ASJC Scopus subject areas

  • Oncology
  • Cancer Research

Cite this

A systems biology approach identifies SART1 as a novel determinant of both 5-fluorouracil and SN38 drug resistance in colorectal cancer. / Allen, Wendy L.; Stevenson, Leanne; Coyle, Vicky M.; Jithesh, Puthen V.; Proutski, Irina; Carson, Gail; Gordon, Michael A.; Lenz, Heinz Josef D; Van Schaeybroeck, Sandra; Longley, Daniel B.; Johnston, Patrick G.

In: Molecular Cancer Therapeutics, Vol. 11, No. 1, 01.2012, p. 119-131.

Research output: Contribution to journalArticle

Allen, WL, Stevenson, L, Coyle, VM, Jithesh, PV, Proutski, I, Carson, G, Gordon, MA, Lenz, HJD, Van Schaeybroeck, S, Longley, DB & Johnston, PG 2012, 'A systems biology approach identifies SART1 as a novel determinant of both 5-fluorouracil and SN38 drug resistance in colorectal cancer', Molecular Cancer Therapeutics, vol. 11, no. 1, pp. 119-131. https://doi.org/10.1158/1535-7163.MCT-11-0510
Allen, Wendy L. ; Stevenson, Leanne ; Coyle, Vicky M. ; Jithesh, Puthen V. ; Proutski, Irina ; Carson, Gail ; Gordon, Michael A. ; Lenz, Heinz Josef D ; Van Schaeybroeck, Sandra ; Longley, Daniel B. ; Johnston, Patrick G. / A systems biology approach identifies SART1 as a novel determinant of both 5-fluorouracil and SN38 drug resistance in colorectal cancer. In: Molecular Cancer Therapeutics. 2012 ; Vol. 11, No. 1. pp. 119-131.
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