Transcriptomic analysis of the effect of ifosfamide on MDCK cells cultivated in microfluidic biochips.

We investigated the behavior of renal cells cultivated in microfluidic biochips when exposed to 50 μM of ifosfamide, an antineoplastic drug treatment. The microarray analysis revealed that ifosfamide had any effect in Petri conditions. The microfluidic biochips induced an early inflammatory response in the MDCK in the untreated cells. This was attributed to cells adapting to the dynamics and micro environment created by the biochips. This led to modulations in the mitochondria dysfunction pathway, the Nrf-2 and oxidative stress pathways and some related cancer genes. When exposed to 50 μM of ifosfamide, we detected a modulation of the pathways related to the cancer and inflammation in the MDCK cultivated in the biochips via modulation of the ATM, p53, MAP Kinase, Nrf-2 and NFKB signaling. In addition, the genes identified and related proteins affected by the ifosfamide treatment in the biochips such as TXNRD1, HSP40 (DNAJB4 and DNAJB9), HSP70 (HSPA9), p21 (CDKN1A), TP53, IKBalpha (NFKBIA) are reported to be the molecular targets in cancer therapy. We also found that the integrin pathway was perturbed with the ifosfamide treatment. Finally, the MYC proto-oncogene appeared to be a potential bridge between the integrin signaling and the anti-inflammatory response.

[1]  G. Rice,et al.  Association between αvβ6 integrin expression, elevated p42/44 kDa MAPK, and plasminogen‐dependent matrix degradation in ovarian cancer , 2002 .

[2]  E. Leonard,et al.  Integrin-mediated RON Growth Factor Receptor Phosphorylation Requires Tyrosine Kinase Activity of Both the Receptor and c-Src* , 2000, The Journal of Biological Chemistry.

[3]  M. Kazanietz,et al.  Constitutive ERK1/2 Activation in Esophagogastric Rib Bone Marrow Micrometastatic Cells Is MEK-independent* , 2001, The Journal of Biological Chemistry.

[4]  Richard A. Miller,et al.  The thioredoxin reductase/thioredoxin system: Novel redox targets for cancer therapy , 2005, Cancer biology & therapy.

[5]  M. Gekle,et al.  Chloroacetaldehyde- and acrolein-induced death of human proximal tubule cells , 2005, Pediatric Nephrology.

[6]  A. Elsharkawy,et al.  Nuclear factor‐κB and the hepatic inflammation‐fibrosis‐cancer axis , 2007 .

[7]  Cécile Legallais,et al.  Analysis of transcriptomic and proteomic profiles demonstrates improved Madin–Darby canine kidney cell function in a renal microfluidic biochip , 2012, Biotechnology progress.

[8]  Shaomeng Wang,et al.  Targeting the MDM2-p53 Interaction for Cancer Therapy , 2008, Clinical Cancer Research.

[9]  Yves Grandvalet,et al.  Integrated Proteomic and Transcriptomic Investigation of the Acetaminophen Toxicity in Liver Microfluidic Biochip , 2011, PloS one.

[10]  F. Oesch,et al.  New Hepatocyte In Vitro Systems for Drug Metabolism: Metabolic Capacity and Recommendations for Application in Basic Research and Drug Development, Standard Operation Procedures , 2003, Drug metabolism reviews.

[11]  G. Burckhardt,et al.  New clues for nephrotoxicity induced by ifosfamide: preferential renal uptake via the human organic cation transporter 2. , 2011, Molecular pharmaceutics.

[12]  M. Taub,et al.  Ifosfamide toxicity in cultured proximal renal tubule cells , 2007, Pediatric Nephrology.

[13]  Donna D. Zhang,et al.  Direct interaction between Nrf2 and p21(Cip1/WAF1) upregulates the Nrf2-mediated antioxidant response. , 2009, Molecular cell.

[14]  A. Levine,et al.  The mdm-2 oncogene product forms a complex with the p53 protein and inhibits p53-mediated transactivation , 1992, Cell.

[15]  P. Bachellier,et al.  Inter-laboratory evaluation of the response of primary human hepatocyte cultures to model CYP inducers - a European Centre for Validation of Alternative Methods (ECVAM) - funded pre-validation study. , 2010, Toxicology in vitro : an international journal published in association with BIBRA.

[16]  R. Riley,et al.  Metabolic screening in vitro: metabolic stability, CYP inhibition and induction. , 2004, Drug discovery today. Technologies.

[17]  R. Dickson,et al.  Regulation of the expression of c-Myc by β1 integrins in epithelial cells , 2001, Oncogene.

[18]  C. Georgopoulos,et al.  The T/t common exon of simian virus 40, JC, and BK polyomavirus T antigens can functionally replace the J-domain of the Escherichia coli DnaJ molecular chaperone. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[19]  T. Yeatman,et al.  Transcriptional recapitulation and subversion of embryonic colon development by mouse colon tumor models and human colon cancer , 2007, Genome Biology.

[20]  Laurent Griscom,et al.  Behavior of HepG2/C3A cell cultures in a microfluidic bioreactor , 2011 .

[21]  T. Xu,et al.  Thioredoxin reductase inactivation as a pivotal mechanism of ifosfamide in cancer therapy. , 2008, European journal of pharmacology.

[22]  I. M. Marks,et al.  The amino-terminal transforming region of simian virus 40 large T and small t antigens functions as a J domain , 1997, Molecular and cellular biology.

[23]  Kou-Juey Wu,et al.  Direct regulation of HSP60 expression by c‐MYC induces transformation , 2008, FEBS letters.

[24]  H. Saito,et al.  Constitutive activation of mitogen-activated protein kinase pathway in acute leukemia cells , 1997, Leukemia.

[25]  Laurent Griscom,et al.  Improvement of HepG2/C3a cell functions in a microfluidic biochip , 2011, Biotechnology and bioengineering.

[26]  P. Workman,et al.  HSP90 as a new therapeutic target for cancer therapy: the story unfolds , 2002, Expert opinion on biological therapy.

[27]  P. Nguyen,et al.  Thioredoxin reductase as a novel molecular target for cancer therapy. , 2006, Cancer letters.

[28]  Roberts Sa,et al.  High-throughput screening approaches for investigating drug metabolism and pharmacokinetics. , 2001 .

[29]  Laurent Griscom,et al.  Development of a Renal Microchip for In Vitro Distal Tubule Models , 2007, Biotechnology progress.

[30]  R. Morimoto,et al.  Role of the heat shock response and molecular chaperones in oncogenesis and cell death. , 2000, Journal of the National Cancer Institute.

[31]  M. Yaffe,et al.  The combined status of ATM and p53 link tumor development with therapeutic response. , 2009, Genes & development.

[32]  O Yoshida,et al.  Constitutive activation of mitogen-activated protein (MAP) kinases in human renal cell carcinoma. , 1995, Cancer research.

[33]  T. Greiner,et al.  Decreased Mdm2 expression inhibits tumor development induced by loss of ARF , 2006, Oncogene.

[34]  Susumu Goto,et al.  KEGG for representation and analysis of molecular networks involving diseases and drugs , 2009, Nucleic Acids Res..

[35]  M. Karin.  The IκB kinase – a bridge between inflammation and cancer , 2008, Cell Research.

[36]  S. Sleijfer,et al.  Decreased exposure to sunitinib due to concomitant administration of ifosfamide: results of a phase I and pharmacokinetic study on the combination of sunitinib and ifosfamide in patients with advanced solid malignancies , 2010, British Journal of Cancer.

[37]  G. Koren,et al.  Ifosfamide nephrotoxicity in children: a mechanistic base for pharmacological prevention , 2009, Expert opinion on drug safety.

[38]  A. Levine,et al.  Mapping of the p53 and mdm-2 interaction domains. , 1993, Molecular and cellular biology.

[39]  P. Silver,et al.  DnaJ/hsp40 chaperone domain of SV40 large T antigen promotes efficient viral DNA replication. , 1997, Genes & development.

[40]  A. Aplin,et al.  Integrin-Mediated Adhesion Regulates ERK Nuclear Translocation and Phosphorylation of Elk-1 , 2001, The Journal of cell biology.

[41]  J. Hahn,et al.  Constitutive activation of extracellular signal-regulated kinase in human acute leukemias: combined role of activation of MEK, hyperexpression of extracellular signal-regulated kinase, and downregulation of a phosphatase, PAC1. , 1999, Blood.