Multifaceted Mechanisms of Cisplatin Resistance in Long-Term Treated Urothelial Carcinoma Cell Lines

Margaretha A. Skowron, Margarita Melnikova, Joep G. H. van Roermund, Andrea Romano, Peter Albers, Juergen Thomale, Wolfgang A. Schulz, Guenter Niegisch, Michele J. Hoffmann*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

13 Citations (Web of Science)

Abstract

Therapeutic efficacy of cisplatin-based treatment of late stage urothelial carcinoma (UC) is limited by chemoresistance. To elucidate underlying mechanisms and to develop new approaches for overcoming resistance, we generated long-term cisplatin treated (LTT) UC cell lines, characterised their cisplatin response, and determined the expression of molecules involved in cisplatin transport and detoxification, DNA repair, and apoptosis. Inhibitors of metallothioneins and Survivin were applied to investigate their ability to sensitise towards cisplatin. Cell growth, proliferation, and clonogenicity were examined after cisplatin treatment by MTT 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, EdU (5-ethynyl-2'-deoxyuridine) incorporation assay, and Giemsa staining, respectively. Cell cycle distribution and apoptosis were quantified by flow cytometry. mRNA and protein expressions were measured by real-time quantitative (qRT)-PCR, western blot, or immunofluorescence staining. LTTs recovered rapidly from cisplatin stress compared to parental cells. In LTTs, to various extents, cisplatin exporters and metallothioneins were induced, cisplatin adduct levels and DNA damage were decreased, whereas expression of DNA repair factors and specific anti-apoptotic factors was elevated. Pharmacological inhibition of Survivin, but not of metallothioneins, sensitised LTTs to cisplatin, in an additive manner. LTTs minimise cisplatin-induced DNA damage and evade apoptosis by increased expression of anti-apoptotic factors. The observed diversity among the four LTTs highlights the complexity of cisplatin resistance mechanisms even within one tumour entity, explaining heterogeneity in patient responses to chemotherapy.
Original languageEnglish
Article number590
Number of pages17
JournalInternational journal of molecular sciences
Volume19
Issue number2
DOIs
Publication statusPublished - 1 Feb 2018

Keywords

  • urothelial carcinoma
  • cisplatin
  • resistance mechanisms
  • metallothionein
  • Survivin
  • YM155
  • CANCER STEM-CELLS
  • BLADDER-CANCER
  • DNA-REPAIR
  • MOLECULAR CHARACTERIZATION
  • ENHANCED EXPRESSION
  • INDUCED APOPTOSIS
  • SURVIVIN
  • METALLOTHIONEIN
  • BIOMARKER
  • CHEMOTHERAPY

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