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Three-dimensional and co-culture models for preclinical evaluation of metal-based anticancer drugs

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Summary

Background Hypoxic and necrotic regions that accrue within solid tumors in vivo are known to be associated with metastasis formation, radio- and chemotherapy resistance, and drug metabolism. Therefore, integration of these tumor characteristics into in vitro drug screening models is advantageous for any reliable investigation of the anticancer activity of novel drug candidates. In general, usage of cell culture models with in vivo like characteristics has become essential in preclinical drug studies and allows evaluation of complex problems such as tumor selectivity and anti-invasive properties of the drug candidates. Materials and Methods In this study, we investigated the anticancer activity of clinically approved, investigational and experimental drugs based on platinum (cisplatin, oxaliplatin and KP1537), gallium (KP46), ruthenium (KP1339) and lanthanum (KP772) in different cell culture models such as monolayers, multicellular spheroids, as well as invasion and metastasis models. Results Application of the Alamar Blue assay to multicellular spheroids and a spheroid-based invasion assay resulted in an altered rating of compounds with regard to their cytotoxicity and ability to inhibit invasion when compared with monolayer-based cytotoxicity and transwell assays. For example, the gallium-based drug candidate KP46 showed in spheroid cultures significantly enhanced properties to inhibit protrusion formation and fibroblast mediated invasiveness, and improved cancer cell selectivity. Conclusion Taken together, our results demonstrate the advantages of spheroid-based assays and underline the necessity of using different experimental models for reliable preclinical investigations assessing and better predicting the anticancer potential of new compounds.

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Abbreviations

CAFs:

Carcinoma associated fibroblasts

CC3:

Caspase 3

CDDP:

Cisplatin

CLSM:

Confocal laser scanning microscopy

DAPI:

4′,6-diamidino-2-phenylindole

DMSO:

Dimethyl sulfoxide

EDTA:

Ethylenediaminetetraacetic acid

FCS:

Fetal calf serum

FGM:

Fibroblast growth medium

GAPDH:

Glycerinaldehyd-3-phosphat-dehydrogenase

HFS:

Hypotonic fluorochrome solution

HIF1α:

Hypoxia-inducible factor 1-alpha

HRP:

Horseradish peroxidase

IC50:

Half maximal inhibitory concentration

I-OHP:

Oxaliplatin

MCR:

Multicellular resistance

MEM:

Minimum essential medium

PBS:

Phosphate buffered saline

PBST:

Phosphate buffered saline with triton X-100

PI:

Propidium iodide

α-SMA:

Alpha-smooth muscle actin

STR:

Short tandem repeat analysis

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Conflict of interest

The authors declare that they have no conflict of interest.

Authorship contributions

Participated in research design: Ekaterina Schreiber-Brynzak, Simone Göschl, Robert Trondl, Helmut Dolznig, Michael A. Jakupec, Bernhard K. Keppler

Conducted experiments: Ekaterina Schreiber-Brynzak, Erik Klapproth, Christine Unger, Irene Lichtscheidl-Schultz, Sarah Schweighofer

Performed data analysis: Ekaterina Schreiber-Brynzak, Erik Klapproth, Simone Göschl, Christine Unger

Wrote or contributed to the writing of the manuscript: Ekaterina Schreiber-Brynzak, Christine Unger, Simone Göschl, Michael A. Jakupec

Author information

Correspondence to Michael A. Jakupec.

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Schreiber-Brynzak, E., Klapproth, E., Unger, C. et al. Three-dimensional and co-culture models for preclinical evaluation of metal-based anticancer drugs. Invest New Drugs 33, 835–847 (2015) doi:10.1007/s10637-015-0260-4

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Keywords

  • 3D models
  • Metal-based drugs
  • Anti-invasive properties
  • Multicellular spheroids
  • Hypoxia