Blockade of SDF-1/CXCR4 signalling inhibits pancreatic cancer progression in vitro via inactivation of canonical Wnt pathway

Extra-pancreatic metastasis is a difficult problem for surgical intervention in pancreatic cancer. CXC chemokine receptor 4 (CXCR4) was considered to have an important role in this process. We hypothesized it may contribute to the pancreatic cancer progression through influencing canonical Wnt pathway. The purpose of this study was to examine the functional role of CXCR4 in the progression of pancreatic cancers and explore the possible mechanism. To this end, the relation between CXCR4 and clinical characteristics was analysed. shRNA against CXCR4 was applied to disrupt the SDF-1/CXCR4 signal transduction pathways in pancreatic cancer cell lines. Our results showed that overall survival in the case of patients positive for CXCR4 expression was significantly lower than that in the case of patients negative for CXCR4 expression. Notably, in vitro studies we observed that the abrogation of CXCR4 could obviously influence the pancreatic cancer cell phenotype including cell proliferation, colony formation, cell invasion and also inhibit the TOPflash activity. In addition, Wnt target genes and mesenchymal markers such as Vimentin and Slug were also inhibited in CXCR4 knockdown cells. Collectively, these data reported here demonstrate CXCR4 could modulate the canonical Wnt pathway and perhaps be a promising therapeutic target for pancreatic cancer progression.

[1]  R. Hruban,et al.  Differentiating pancreatic lesions by microarray and QPCR analysis of pancreatic juice RNAs , 2006, Cancer biology & therapy.

[2]  T. Mcclanahan,et al.  Involvement of chemokine receptors in breast cancer metastasis , 2001, Nature.

[3]  R. Nawroth,et al.  Extracellular Sulfatases, Elements of the Wnt Signaling Pathway, Positively Regulate Growth and Tumorigenicity of Human Pancreatic Cancer Cells , 2007, PloS one.

[4]  Xiao-Hua Wu,et al.  Expression of chemokine CXCL12 and its receptor CXCR4 in human epithelial ovarian cancer: an independent prognostic factor for tumor progression. , 2006, Gynecologic oncology.

[5]  M. Probst-Kepper,et al.  CXCR4/CXCL12 expression and signalling in kidney cancer , 2002, British Journal of Cancer.

[6]  H. Clevers,et al.  Wnt signalling in stem cells and cancer , 2005, Nature.

[7]  P. Galle,et al.  Strong expression of chemokine receptor CXCR4 by pancreatic cancer correlates with advanced disease. , 2006, Oncology reports.

[8]  V. Shah,et al.  Characterization of the CXCR4 Signaling in Pancreatic Cancer Cells , 2007, International journal of gastrointestinal cancer.

[9]  P. Murphy,et al.  Chemokines and the molecular basis of cancer metastasis. , 2001, The New England journal of medicine.

[10]  Michael A. Choti,et al.  1423 pancreaticoduodenectomies for pancreatic cancer: A single-institution experience , 2006, Journal of Gastrointestinal Surgery.

[11]  Thomas D. Schmittgen,et al.  Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method. , 2001, Methods.

[12]  Tracy M Handel,et al.  Chemokines and cancer: migration, intracellular signalling and intercellular communication in the microenvironment. , 2008, The Biochemical journal.

[13]  J. Kench,et al.  Common Activation of Canonical Wnt Signaling in Pancreatic Adenocarcinoma , 2007, PloS one.

[14]  R. Knuechel,et al.  Variable β-catenin expression in colorectal cancers indicates tumor progression driven by the tumor environment , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[15]  N. Warrington,et al.  Widespread CXCR4 activation in astrocytomas revealed by phospho-CXCR4-specific antibodies. , 2005, Cancer research.

[16]  M. Locati,et al.  Hepatocyte growth factor enhances CXCR4 expression favoring breast cancer cell invasiveness. , 2005, Experimental cell research.

[17]  S. Brand,et al.  CXCR4 and CXCL12 are inversely expressed in colorectal cancer cells and modulate cancer cell migration, invasion and MMP-9 activation. , 2005, Experimental cell research.

[18]  M. Tsao,et al.  Epithelial-Cadherin and β-Catenin Expression Changes in Pancreatic Intraepithelial Neoplasia , 2004, Clinical Cancer Research.

[19]  A. Bell,et al.  Aberrant Wnt/beta-catenin signaling in pancreatic adenocarcinoma. , 2006, Neoplasia.

[20]  A. Okada,et al.  Mutation of beta-catenin and its protein accumulation in solid and cystic tumor of the pancreas associated with metastasis. , 2003, International journal of molecular medicine.

[21]  Fumitaka Kikkawa,et al.  Involvement of SDF-1alpha/CXCR4 axis in the enhanced peritoneal metastasis of epithelial ovarian carcinoma. , 2008, International journal of cancer.

[22]  G. Botti,et al.  Inhibitory effects of anti-CXCR4 antibodies on human colon cancer cells , 2005, Cancer Immunology, Immunotherapy.

[23]  M. Schwaiger,et al.  CXCR4 expression increases liver and lung metastasis in a mouse model of pancreatic cancer. , 2005, Gastroenterology.

[24]  Slug expression is an independent prognostic parameter for poor survival in colorectal carcinoma patients , 2006, British Journal of Cancer.

[25]  M. Ilyas,et al.  Wnt signalling and the mechanistic basis of tumour development , 2005, The Journal of pathology.

[26]  N. Dubrawsky.  Cancer statistics , 1989, CA: a cancer journal for clinicians.

[27]  T. Kipps,et al.  CXCR4: a key receptor in the crosstalk between tumor cells and their microenvironment. , 2006, Blood.

[28]  H. Yoshida,et al.  Epithelial-mesenchymal transition induced by the stromal cell-derived factor-1/CXCR4 system in oral squamous cell carcinoma cells. , 2006, International journal of oncology.

[29]  R. Nusse,et al.  Convergence of Wnt, ß-Catenin, and Cadherin Pathways , 2004, Science.

[30]  N. Clarke,et al.  Invasive characteristics of human prostatic epithelial cells: understanding the metastatic process , 2005, British Journal of Cancer.

[31]  H. Nakshatri,et al.  NF-κ B Promotes Breast Cancer Cell Migration and Metastasis by Inducing the Expression of the Chemokine Receptor CXCR4* , 2003, Journal of Biological Chemistry.

[32]  M. Taketo.  Wnt signaling and gastrointestinal tumorigenesis in mouse models , 2006, Oncogene.

[33]  H. Moch,et al.  Chemokine receptor CXCR4 downregulated by von Hippel–Lindau tumour suppressor pVHL , 2003, Nature.

[34]  F. Balkwill.  The significance of cancer cell expression of the chemokine receptor CXCR4. , 2004, Seminars in cancer biology.

[35]  R. Nusse,et al.  Convergence of Wnt, beta-catenin, and cadherin pathways. , 2004, Science.

[36]  M. Menger,et al.  Stromal cell-derived factor-1 promotes cell migration and tumor growth of colorectal metastasis. , 2007, Neoplasia.

[37]  Fumitaka Kikkawa,et al.  Involvement of SDF‐1α/CXCR4 axis in the enhanced peritoneal metastasis of epithelial ovarian carcinoma , 2008 .

[38]  C. Fenoglio-Preiser,et al.  Dysregulation of β-Catenin Expression Correlates With Tumor Differentiation in Pancreatic Duct Adenocarcinoma , 2003, Annals of Surgical Oncology.

[39]  F. Balkwill.  Cancer and the chemokine network , 2004, Nature Reviews Cancer.

[40]  G. Hildebrandt,et al.  Expression of Erythropoietin and Erythropoietin Receptor in Cervical Cancer and Relationship to Survival, Hypoxia, and Apoptosis , 2006, Clinical Cancer Research.

[41]  J. Cameron,et al.  Solid-pseudopapillary tumors of the pancreas are genetically distinct from pancreatic ductal adenocarcinomas and almost always harbor beta-catenin mutations. , 2002, The American journal of pathology.

[42]  T. Takada,et al.  Influence of resection margins and treatment on survival in patients with pancreatic cancer: meta-analysis of randomized controlled trials. , 2008, Archives of surgery.

[43]  J. Votaw,et al.  Silencing of CXCR4 blocks breast cancer metastasis. , 2005, Cancer research.

[44]  P. Neth,et al.  The Wnt Signal Transduction Pathway in Stem Cells and Cancer Cells: Influence on Cellular Invasion , 2007, Stem Cell Reviews.

[45]  Q. Ma,et al.  β-Catenin is a promising key factor in the SDF-1/CXCR4 axis on metastasis of pancreatic cancer☆ , 2007 .

[46]  A. Jemal,et al.  Cancer Statistics, 2007 , 2007, CA: a cancer journal for clinicians.

[47]  M. Peifer,et al.  Wnt signaling in oncogenesis and embryogenesis--a look outside the nucleus. , 2000, Science.

[48]  J. Thiery.  Epithelial–mesenchymal transitions in tumour progression , 2002, Nature Reviews Cancer.

[49]  A. Bell,et al.  Aberrant Wnt/β-Catenin Signaling in Pancreatic Adenocarcinoma , 2006 .

[50]  J. Imura,et al.  Expression of SDF-1α and nuclear CXCR4 predicts lymph node metastasis in colorectal cancer , 2008, British Journal of Cancer.

[51]  M. Schuler,et al.  Dissemination of hepatocellular carcinoma is mediated via chemokine receptor CXCR4 , 2006, British Journal of Cancer.

[52]  Z. Wang,et al.  Beta-catenin is a promising key factor in the SDF-1/CXCR4 axis on metastasis of pancreatic cancer. , 2007, Medical Hypotheses.

[53]  A. Sadanandam,et al.  Chemokines in tumor angiogenesis and metastasis , 2007, Cancer and Metastasis Reviews.

[54]  R. Bachelder,et al.  Vascular endothelial growth factor promotes breast carcinoma invasion in an autocrine manner by regulating the chemokine receptor CXCR4. , 2002, Cancer research.