title: Real-time monitoring of cisplatin-induced cell death.
abstract: Since the discovery of cisplatin more than 40 years ago and its clinical introduction in the 1970s an enormous amount of research has gone into elucidating the mechanism of action of cisplatin on tumor cells. With a novel cell biosensor chip system allowing continuous monitoring of respiration, glycolysis, and impedance we followed cisplatin treatment of different cancer cell lines in real-time. Our measurements reveal a first effect on respiration, in all cisplatin treated cell lines, followed with a significant delay by interference with glycolysis in HT-29, HCT-116, HepG2, and MCF-7 cells but not in the cisplatin-resistant cell line MDA-MB-231. Most strikingly, cell death started in all cisplatin-sensitive cell lines within 8 to 11 h of treatment, indicating a clear time frame from exposure, first response to cisplatin lesions, to cell fate decision. The time points of most significant changes were selected for more detailed analysis of cisplatin response in the breast cancer cell line MCF-7. Phosphorylation of selected signal transduction mediators connected with cellular proliferation, as well as changes in gene expression, were analyzed in samples obtained directly from sensor chips at the time points when changes in glycolysis and impedance occurred. Our online cell biosensor measurements reveal for the first time the time scale of metabolic response until onset of cell death under cisplatin treatment, which is in good agreement with models of p53-mediated cell fate decision.
authors: Institute of Pharmacy and Molecular Biotechnology, Heidelberg University, Heidelberg, Germany.
authors: Hamed
authors: H
authors: Alborzinia
authors: Suzan
authors: S
authors: Can
authors: Pavlo
authors: P
authors: Holenya
authors: Catharina
authors: C
authors: Scholl
authors: Elke
authors: E
authors: Lederer
authors: Igor
authors: I
authors: Kitanovic
authors: Stefan
authors: S
authors: Wölfl
references: J Bioenerg Biomembr. 2007 Feb;39(1):43-50
references: 17318397
references: Oncogene. 2003 Nov 6;22(50):8168-77
references: 14603257
references: Nucleic Acids Res. 2002 Jun 1;30(11):e50
references: 12034851
references: Mol Cancer Ther. 2009 Jul;8(7):1916-23
references: 19567816
references: J Biol Chem. 2000 Dec 15;275(50):39435-43
references: 10993883
references: Genome Biol. 2003;4(5):P3
references: 12734009
references: Cancer Treat Rep. 1976 Jan;60(1):55-60
references: 1000519
references: J Med Chem. 2010 Dec 23;53(24):8608-18
references: 21082862
references: Proc Natl Acad Sci U S A. 2001 Jan 2;98(1):31-6
references: 11134512
references: Biophys J. 2010 Sep 22;99(6):1696-705
references: 20858413
references: Biochem J. 2008 Aug 15;414(1):93-102
references: 18426391
references: Nature. 2000 Nov 16;408(6810):307-10
references: 11099028
references: Genes Dev. 1999 Nov 15;13(22):2905-27
references: 10579998
references: J Appl Toxicol. 2005 Sep-Oct;25(5):374-82
references: 16013042
references: Fresenius J Anal Chem. 2001 Jan 1;369(1):30-5
references: 11210226
references: J Biol Chem. 2002 Jan 11;277(2):1255-60
references: 11705991
references: J Med Chem. 2008 Jul 10;51(13):3924-33
references: 18543901
references: Exp Cell Res. 2000 Aug 25;259(1):158-66
references: 10942588
references: Nat Protoc. 2009;4(1):44-57
references: 19131956
references: J Biol Chem. 2008 Jan 11;283(2):726-32
references: 17991738
references: Crit Rev Oncol Hematol. 2005 Jan;53(1):3-11
references: 15607931
references: Proc Natl Acad Sci U S A. 1991 Sep 1;88(17):7896-900
references: 1881923
references: Proteomics. 2011 May;11(10):2129-33
references: 21500346
references: Nature. 1965 Feb 13;205:698-9
references: 14287410
references: J Biol Chem. 2008 Feb 15;283(7):3731-42
references: 18079115
references: Nat Rev Drug Discov. 2005 Apr;4(4):307-20
references: 15789122