Platinum complex cytotoxicity tested by the electrical resistance breakdown assay.
The electrical resistance breakdown assay provides a novel approach for the quantification of cytotoxic activity of platinum based anticancer drugs. It is a functional assay system for cancer cell invasion that detects nanoscale alterations of an epithelial test barrier prior to microscopic morphometric changes. We measured changes in transepithelial electrical resistance (TEER) of a tight epithelial MDCK-C7 monolayer in response to highly invasive amelanotic melanoma cells (A7-clone) in combination with different platinum complexes (cis-, oxali- and carboplatin). The efficiency of the electrical resistance breakdown assay was compared a standard method for measurement of cytostatic activity, the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay. The MTT-assay utilizes mitochondrial enzymatic activity to draw conclusions from a functional cell metabolism to the number of living cells in a sample. When human melanoma cells were seeded on top of an electrically tight MDCK-C7 monolayer, electrical leakage occurred within 48 h of co-culture. Electrical resistance breakdown was effectively prevented by cisplatin and its analogs (no significant difference between 100 microM cisplatin and corresponding controls with non-invasive cells). The results of the electrical resistance breakdown and MTT-assay were linearly dependent. Significance of both tests was equivalent, but the electrical resistance breakdown assay gave additional functional information. Compared to oxali- and carboplatin, cisplatin was more effective in preventing TEER-breakdown than reducing the number of tumor cells, giving rise to the assumption that cisplatin can reduce tumor cell number as well as invasiveness. In conclusion the electrical resistance breakdown assay provides a sensitive, continuous and cell-based assay system for the quantification of cancer cell invasiveness and evaluation of chemotherapeutics under physiological conditions.
Ludwig
Thomas
T
Institute of Physiology II, University of Münster, Germany. thomas.ludwig@uni-muenster.de
Fakih
Sarah
S
Krebs
Bernt
B
Oberleithner
Hans
H
D000818:Animals; D000970:Antineoplastic Agents / Q000633:toxicity*; D001681:Biological Assay / Q000295:instrumentation; D045744:Cell Line, Tumor; D004285:Dogs; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D017097:Electric Impedance; D004847:Epithelial Cells / Q000187:drug effects / Q000502:physiology; D006801:Humans; D018328:Melanoma, Amelanotic / Q000737:chemistry / Q000378:metabolism; D017671:Platinum Compounds / Q000633:toxicity*
D000970:Antineoplastic Agents; D017671:Platinum Compounds