Molecular characterization of irinotecan (SN-38) resistant human breast cancer cell lines

BMC Cancer. 2016 Jan 22:16:34. doi: 10.1186/s12885-016-2071-1.

Abstract

Background: Studies in taxane and/or anthracycline refractory metastatic breast cancer (mBC) patients have shown approximately 30% response rates to irinotecan. Hence, a significant number of patients will experience irinotecan-induced side effects without obtaining any benefit. The aim of this study was to lay the groundwork for development of predictive biomarkers for irinotecan treatment in BC.

Methods: We established BC cell lines with acquired or de novo resistance to SN-38, by exposing the human BC cell lines MCF-7 and MDA-MB-231 to either stepwise increasing concentrations over 6 months or an initial high dose of SN-38 (the active metabolite of irinotecan), respectively. The resistant cell lines were analyzed for cross-resistance to other anti-cancer drugs, global gene expression, growth rates, TOP1 and TOP2A gene copy numbers and protein expression, and inhibition of the breast cancer resistance protein (ABCG2/BCRP) drug efflux pump.

Results: We found that the resistant cell lines showed 7-100 fold increased resistance to SN-38 but remained sensitive to docetaxel and the non-camptothecin Top1 inhibitor LMP400. The resistant cell lines were characterized by Top1 down-regulation, changed isoelectric points of Top1 and reduced growth rates. The gene and protein expression of ABCG2/BCRP was up-regulated in the resistant sub-lines and functional assays revealed BCRP as a key mediator of SN-38 resistance.

Conclusions: Based on our preclinical results, we suggest analyzing the predictive value of the BCRP in breast cancer patients scheduled for irinotecan treatment. Moreover, LMP400 should be tested in a clinical setting in breast cancer patients with resistance to irinotecan.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters / biosynthesis
  • ATP-Binding Cassette Transporters / genetics*
  • Antigens, Neoplasm / genetics
  • Breast Neoplasms / drug therapy*
  • Breast Neoplasms / genetics*
  • Breast Neoplasms / pathology
  • Camptothecin / administration & dosage
  • Camptothecin / adverse effects
  • Camptothecin / analogs & derivatives*
  • DNA Topoisomerases, Type I / biosynthesis
  • DNA Topoisomerases, Type I / genetics*
  • DNA Topoisomerases, Type II / genetics
  • DNA-Binding Proteins / genetics
  • Docetaxel
  • Drug Resistance, Neoplasm / genetics
  • Female
  • Gene Dosage / genetics
  • Gene Expression Regulation, Neoplastic / drug effects
  • Humans
  • Irinotecan
  • MCF-7 Cells
  • Neoplasm Proteins / biosynthesis
  • Neoplasm Proteins / genetics*
  • Poly-ADP-Ribose Binding Proteins
  • Taxoids / administration & dosage

Substances

  • ABCG2 protein, human
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • ATP-Binding Cassette Transporters
  • Antigens, Neoplasm
  • DNA-Binding Proteins
  • Neoplasm Proteins
  • Poly-ADP-Ribose Binding Proteins
  • Taxoids
  • Docetaxel
  • Irinotecan
  • DNA Topoisomerases, Type I
  • TOP1 protein, human
  • DNA Topoisomerases, Type II
  • TOP2A protein, human
  • Camptothecin