Microarray-Assisted Pathway Analysis Identifies MT1X & NFκB as Mediators of TCRP1-Associated Resistance to Cisplatin in Oral Squamous Cell Carcinoma

We recently reported that TCRP1, a novel multidrug-resistance associated human gene, can mediate cisplatin resistance in OSCC cells. However, the molecular mechanism underlying this role of TCRP1 remained to be elucidated. In this study, by using Human Toxicology and Drug Resistance Microarray, we identified 30 genes with significantly different expression levels between Tca/PYM and TCRP1 knockdown cell lines. Co-immunoprecipitation experiments and GST-pull down assays showed that metallothionein1X (MT1X) and Akt interact with TCRP1. siRNA-mediated knockdown of TCRP1 and MT1X was found to sensitize cells to cisplatin, leading to increased apoptosis and inhibition of cell proliferation. These functions of TCRP1 may be caused at least in part via activation of the PI3K/Akt/NF-κB signaling pathway. Taken together, our findings indicate that TCRP1 may be an important drug target for improvement of the treatment and survival of patients with oral squamous cell carcinoma.

[1]  Zhuchu Chen,et al.  High-yield expression in Escherichia coli of soluble human MT2A with native functions. , 2007, Protein expression and purification.

[2]  V. B. Konkimalla,et al.  Role of transporter genes in cisplatin resistance. , 2008, In vivo.

[3]  R. Altman,et al.  Platinum pathway. , 2009, Pharmacogenetics and genomics.

[4]  J. Essigmann,et al.  Mechanisms of resistance to cisplatin. , 2001, Mutation research.

[5]  Y. Xiong,et al.  Cloning and functional characterization of TCRP1, a novel gene mediating resistance to cisplatin in an oral squamous cell carcinoma cell line , 2011, FEBS letters.

[6]  Brendan M Leung,et al.  p53 is a determinant of X-linked inhibitor of apoptosis protein/Akt-mediated chemoresistance in human ovarian cancer cells. , 2003, Cancer research.

[7]  José Perez,et al.  Is cisplatin-induced cell death always produced by apoptosis? , 2001, Molecular pharmacology.

[8]  W. ElShamy,et al.  BRCA1-IRIS overexpression promotes cisplatin resistance in ovarian cancer cells. , 2010, Cancer research.

[9]  G. Chu,et al.  Cellular responses to cisplatin. The roles of DNA-binding proteins and DNA repair. , 1994, The Journal of biological chemistry.

[10]  Yi-xue Gu,et al.  TCRP1 promotes radioresistance of oral squamous cell carcinoma cells via Akt signal pathway , 2011, Molecular and Cellular Biochemistry.

[11]  Il-Jin Kim,et al.  Identification of Genes with Differential Expression in Acquired Drug-Resistant Gastric Cancer Cells Using High-Density Oligonucleotide Microarrays , 2004, Clinical Cancer Research.

[12]  R. Cousins,et al.  Zinc supplementation of young men alters metallothionein, zinc transporter, and cytokine gene expression in leukocyte populations. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[13]  Biaoyang Lin,et al.  Proteins Associated with Cisplatin Resistance in Ovarian Cancer Cells Identified by Quantitative Proteomic Technology and Integrated with mRNA Expression Levels*S , 2006, Molecular & Cellular Proteomics.

[14]  Jong-Ho Lee,et al.  The role of tamoxifen in combination with cisplatin on oral squamous cell carcinoma cell lines. , 2007, Cancer letters.

[15]  J. Bourhis,et al.  Chemotherapy added to locoregional treatment for head and neck squamous-cell carcinoma: three meta-analyses of updated individual data , 2000, The Lancet.

[16]  P. Jordan,et al.  Molecular mechanisms involved in cisplatin cytotoxicity , 2000, Cellular and Molecular Life Sciences CMLS.

[17]  William C Reinhold,et al.  Membrane Transporters and Channels , 2004, Cancer Research.

[18]  Yung-Hyun Choi,et al.  Pectenotoxin-2 abolishes constitutively activated NF-kappaB, leading to suppression of NF-kappaB related gene products and potentiation of apoptosis. , 2008, Cancer letters.

[19]  Hyung Gyun Kim,et al.  Metallothionein-III provides neuronal protection through activation of nuclear factor-kappaB via the TrkA/phosphatidylinositol-3 kinase/Akt signaling pathway. , 2009, Toxicological sciences : an official journal of the Society of Toxicology.

[20]  J. Pignon,et al.  Chemotherapy added to locoregional treatment for head and neck squamous-cell carcinoma: three meta-analyses of updated individual data. MACH-NC Collaborative Group. Meta analysis of chemotherapy on head and neck cancer , 2001 .

[21]  J. Groten,et al.  Role of metallothionein in cisplatin sensitivity of germ‐cell tumours , 2000, International journal of cancer.

[22]  Dong M Shin,et al.  Recent advances in head and neck cancer. , 2008, The New England journal of medicine.

[23]  P. Moynagh,et al.  The NF-kappaB pathway. , 2005, Journal of cell science.

[24]  O. Fardel,et al.  Blockage of Multidrug Resistance-Associated Proteins Potentiates the Inhibitory Effects of Arsenic Trioxide on CYP1A1 Induction by Polycyclic Aromatic Hydrocarbons , 2003, Journal of Pharmacology and Experimental Therapeutics.

[25]  Howard L McLeod,et al.  PI3K/Akt/mTOR pathway as a target for cancer therapy , 2005, Anti-cancer drugs.

[26]  Yi-xue Gu,et al.  Purification and biochemical characterization of a novel protein-tongue cancer chemotherapy resistance-associated protein1 (TCRP1). , 2012, Protein expression and purification.

[27]  S. Bates,et al.  Effect of Common CYP3A4 and CYP3A5 Variants on the Pharmacokinetics of the Cytochrome P450 3A Phenotyping Probe Midazolam in Cancer Patients , 2005, Clinical Cancer Research.

[28]  Ping Zhang,et al.  Identification of genes associated with cisplatin resistance in human oral squamous cell carcinoma cell line , 2006, BMC Cancer.

[29]  P. Moynagh,et al.  The NF-κB pathway , 2005, Journal of Cell Science.

[30]  G. Kong,et al.  Id-1 activates Akt-mediated Wnt signaling and p27Kip1 phosphorylation through PTEN inhibition , 2009, Oncogene.

[31]  Hyeong-Seok Lim,et al.  Increased Expression of Metallothionein Is Associated with Irinotecan Resistance in Gastric Cancer , 2004, Cancer Research.

[32]  Y. Xiong,et al.  14‐3‐3σ regulation by p53 mediates a chemotherapy response to 5‐fluorouracil in MCF‐7 breast cancer cells via Akt inactivation , 2012, FEBS letters.

[33]  N. Imura,et al.  Modulation of resistance to anticancer drugs by inhibition of metallothionein synthesis. , 1994, Cancer research.

[34]  Zhiming He,et al.  Identification of carbonic anhydrase 9 as a contributor to pingyangmycin‐induced drug resistance in human tongue cancer cells , 2010, The FEBS journal.

[35]  T. Hudson,et al.  Disruption of AP1S1, Causing a Novel Neurocutaneous Syndrome, Perturbs Development of the Skin and Spinal Cord , 2008, PLoS genetics.

[36]  R. Weinshilboum,et al.  Sulfotransferase gene copy number variation: pharmacogenetics and function , 2009, Cytogenetic and Genome Research.

[37]  J. Beijnen,et al.  Characterization of Drug Transport by the Human Multidrug Resistance Protein 3 (ABCC3)* , 2001, The Journal of Biological Chemistry.

[38]  D. Winge.  Limited proteolysis of metallothioneins. , 1991, Methods in enzymology.

[39]  K. Uzawa,et al.  Identification of cisplatin‐resistance related genes in head and neck squamous cell carcinoma , 2010, International journal of cancer.