Myeloid Zinc Finger 1 Induces Migration, Invasion, and In vivo Metastasis through Axl Gene Expression in Solid Cancer

Myeloid zinc finger 1 (MZF1) is a member of the SCAN domain family transcription factors that form dimers through their highly conserved SCAN motifs. Silencing of MZF1 inhibits cell proliferation, and abnormal expression of MZF1 results in cancer development. However, a potential role of MZF1 in metastasis remains unclear. Axl is a receptor tyrosine kinase and was first identified as a transforming gene in chronic myeloid leukemia. Axl overexpression induces proliferation, migration, and invasion and is highly expressed in different human cancers. In this study, we show that overexpression of MZF1 induces migration and invasion in colorectal (Rko, SW480) and cervical (HeLa) cancer cells. In addition, we show that MZF1 binds to the Axl promoter, transactivates promoter activity, and enhances Axl-mRNA and protein expression in a dose-dependent manner. In vitro, sh-RNA knockdown of Axl reduced MZF1-induced migration and invasion in HeLa and Rko cells (P = 0.05). Additionally, Rko cells overexpressing MZF1 showed increased tumor formation and liver metastasis in the chicken-embryo-metastasis assay in vivo. Furthermore, the expression of MZF1 and Axl was significantly higher in resected colorectal tumors compared with corresponding normal tissues (P = 0.02; P = 0.05), and MZF1 expression was positively correlated with Axl gene expression in tumor tissues (P < 0.01). Taken together, this is the first study to show that MZF1 induces invasion and in vivo metastasis in colorectal and cervical cancer, at least in part by regulating Axl gene expression. Mol Cancer Res; 8(2); 159–69.

[1]  E. Liu,et al.  Determinants for transformation induced by the Axl receptor tyrosine kinase , 1998, Oncogene.

[2]  T. Sander,et al.  Characterization of the SCAN box encoding RAZ1 gene: analysis of cDNA transcripts, expression, and cellular localization. , 2002, Gene.

[3]  H. Hanafusa,et al.  Identification of the Product of Growth Arrest-specific Gene 6 as a Common Ligand for Axl, Sky, and Mer Receptor Tyrosine Kinases* , 1996, The Journal of Biological Chemistry.

[4]  E. Liu,et al.  Differential activation of the Ras/extracellular-signal-regulated protein kinase pathway is responsible for the biological consequences induced by the Axl receptor tyrosine kinase , 1996, Molecular and cellular biology.

[5]  A. Ullrich,et al.  Intracellular signaling of the Ufo/Axl receptor tyrosine kinase is mediated mainly by a multi-substrate docking-site , 1997, Oncogene.

[6]  L. Castello,et al.  Gas6 induces proliferation in prostate carcinoma cell lines expressing the Axl receptor , 2005, Journal of cellular physiology.

[7]  S. Tsai,et al.  Parallel hybridization analysis of multiple protein kinase genes: identification of gene expression patterns characteristic of human hepatocellular carcinoma. , 1998, Genomics.

[8]  B Calabretta,et al.  Overexpression of the zinc finger protein MZF1 inhibits hematopoietic development from embryonic stem cells: correlation with negative regulation of CD34 and c-myb promoter activity , 1995, Molecular and cellular biology.

[9]  A. Schulz,et al.  A novel putative tyrosine kinase receptor with oncogenic potential. , 1991, Oncogene.

[10]  J. Yokota,et al.  Biological properties and gene expression associated with metastatic potential of human osteosarcoma , 2004, Clinical & Experimental Metastasis.

[11]  H. Allgayer,et al.  The human receptor tyrosine kinase Axl gene--promoter characterization and regulation of constitutive expression by Sp1, Sp3 and CpG methylation. , 2008, Bioscience reports.

[12]  Lennart Philipson,et al.  Genes specifically expressed at growth arrest of mammalian cells , 1988, Cell.

[13]  Y. Chu,et al.  Expression of axl in lung adenocarcinoma and correlation with tumor progression. , 2005, Neoplasia.

[14]  R. Hromas,et al.  The myeloid zinc finger gene (MZF-1) delays retinoic acid-induced apoptosis and differentiation in myeloid leukemia cells , 1998, Leukemia.

[15]  E. Dreher,et al.  Estrogen dependent expression of the receptor tyrosine kinase axl in normal and malignant human breast. , 2001, Annals of oncology : official journal of the European Society for Medical Oncology.

[16]  E. Ruaro,et al.  Gas6-mediated survival in NIH3T3 cells activates stress signalling cascade and is independent of Ras , 1999, Oncogene.

[17]  M. Klemsz,et al.  The myeloid zinc finger gene, MZF-1, regulates the CD34 promoter in vitro. , 1995, Blood.

[18]  O. Fromigué,et al.  Sp1/Sp3 and the myeloid zinc finger gene MZF1 regulate the human N-cadherin promoter in osteoblasts. , 2005, Experimental cell research.

[19]  C. Basilico,et al.  The receptor tyrosine kinase ARK mediates cell aggregation by homophilic binding , 1995, Molecular and cellular biology.

[20]  P. Pandolfi,et al.  Mzf1 controls cell proliferation and tumorigenesis. , 2001, Genes & development.

[21]  K. Cornetta,et al.  Aberrant expression of the myeloid zinc finger gene, MZF-1, is oncogenic. , 1995, Cancer research.

[22]  S. Goff,et al.  Signaling through the ARK tyrosine kinase receptor protects from apoptosis in the absence of growth stimulation , 1997, Oncogene.

[23]  B. Dahlbäck,et al.  Vitamin K-dependent Gas6 activates ERK kinase and stimulates growth of cardiac fibroblasts. , 2004, Biochemical and biophysical research communications.

[24]  K. Heidenreich,et al.  Adhesion-related kinase induction of migration requires phosphatidylinositol-3-kinase and ras stimulation of rac activity in immortalized gonadotropin-releasing hormone neuronal cells. , 2007, Endocrinology.

[25]  Stephen T. C. Wong,et al.  Multiple roles for the receptor tyrosine kinase axl in tumor formation. , 2005, Cancer research.

[26]  H. Allgayer,et al.  MicroRNA-21 (miR-21) post-transcriptionally downregulates tumor suppressor Pdcd4 and stimulates invasion, intravasation and metastasis in colorectal cancer , 2008, Oncogene.

[27]  R. Hromas,et al.  Antisense oligonucleotides from the stage-specific myeloid zinc finger gene MZF-1 inhibit granulopoiesis in vitro , 1991, The Journal of experimental medicine.

[28]  K. Cornetta,et al.  Forced over-expression of the myeloid zinc finger gene MZF-1 inhibits apoptosis and promotes oncogenesis in interleukin-3-dependent FDCP.1 cells. , 1996, Leukemia.

[29]  D. Tuan,et al.  The Long Terminal Repeat (LTR) of ERV-9 Human Endogenous Retrovirus Binds to NF-Y in the Assembly of an Active LTR Enhancer Complex NF-Y/MZF1/GATA-2* , 2005, Journal of Biological Chemistry.

[30]  Y. Hsieh,et al.  Suppression of tumorigenicity of human hepatocellular carcinoma cells by antisense oligonucleotide MZF-1. , 2007, The Chinese journal of physiology.

[31]  D. Albert,et al.  Expression of the Receptor Tyrosine Kinase Axl Promotes Ocular Melanoma Cell Survival , 2004, Cancer Research.

[32]  A. Kawada,et al.  Crucial roles of MZF1 and Sp1 in the transcriptional regulation of the peptidylarginine deiminase type I gene (PADI1) in human keratinocytes. , 2008, The Journal of investigative dermatology.

[33]  M. Klemsz,et al.  Hematopoietic transcriptional regulation by the myeloid zinc finger gene, MZF-1. , 1996, Current topics in microbiology and immunology.

[34]  R. Herrmann,et al.  Axl expression is associated with adverse prognosis and with expression of Bcl-2 and CD34 in de novo acute myeloid leukemia (AML): results from a multicenter trial of the Swiss Group for Clinical Cancer Research (SAKK) , 1999, Leukemia.

[35]  M. Essig,et al.  Dominant-negative inhibition of the Axl receptor tyrosine kinase suppresses brain tumor cell growth and invasion and prolongs survival. , 2006, Proceedings of the National Academy of Sciences of the United States of America.

[36]  F. Meric,et al.  Expression profile of tyrosine kinases in breast cancer. , 2002, Clinical cancer research : an official journal of the American Association for Cancer Research.

[37]  R. Paus,et al.  A promiscuous liaison between IL‐15 receptor and Axl receptor tyrosine kinase in cell death control , 2005, The EMBO journal.

[38]  B. Berk,et al.  Axl, A Receptor Tyrosine Kinase, Mediates Flow-Induced Vascular Remodeling , 2006, Circulation research.

[39]  E. Kandel,et al.  Novel Mechanism for Gonadotropin-Releasing Hormone Neuronal Migration Involving Gas6/Ark Signaling to p38 Mitogen-Activated Protein Kinase , 2002, Molecular and Cellular Biology.

[40]  C. Chi,et al.  Clinical significance of AXL kinase family in gastric cancer. , 2002, Anticancer research.

[41]  E. Attar,et al.  GAS6 Induces Axl-mediated Chemotaxis of Vascular Smooth Muscle Cells* , 1998, The Journal of Biological Chemistry.

[42]  M. Cavet,et al.  Gas6 inhibits apoptosis in vascular smooth muscle: role of Axl kinase and Akt. , 2004, Journal of molecular and cellular cardiology.

[43]  Yi Guo,et al.  MZF1 possesses a repressively regulatory function in ERCC1 expression. , 2006, Biochemical pharmacology.

[44]  R. Verardo,et al.  Gas6 Anti-apoptotic Signaling Requires NF-κB Activation* , 2001, The Journal of Biological Chemistry.

[45]  G. Mills,et al.  Cetuximab attenuates metastasis and u-PAR expression in non-small cell lung cancer: u-PAR and E-cadherin are novel biomarkers of cetuximab sensitivity. , 2009, Cancer research.

[46]  R. Hromas,et al.  Characterization of the DNA-binding properties of the myeloid zinc finger protein MZF1: two independent DNA-binding domains recognize two DNA consensus sequences with a common G-rich core , 1994, Molecular and cellular biology.

[47]  Michael Joseph Peterson,et al.  Identification of a Novel SCAN Box-related Protein That Interacts with MZF1B , 2000, The Journal of Biological Chemistry.

[48]  Y. Hsieh,et al.  PKCα expression regulated by Elk-1 and MZF-1 in human HCC cells , 2006 .

[49]  E. Liu,et al.  Receptor tyrosine kinases expressed in metastatic colon cancer , 1995, International journal of cancer.

[50]  A. Ullrich,et al.  Axl and Growth Arrest–Specific Gene 6 Are Frequently Overexpressed in Human Gliomas and Predict Poor Prognosis in Patients with Glioblastoma Multiforme , 2008, Clinical Cancer Research.

[51]  B. Varnum,et al.  GAS6 Mediates Adhesion of Cells Expressing the Receptor Tyrosine Kinase Axl* , 1997, The Journal of Biological Chemistry.

[52]  L. Deaven,et al.  A retinoic acid-responsive human zinc finger gene, MZF-1, preferentially expressed in myeloid cells. , 1991, The Journal of biological chemistry.

[53]  H. Allgayer,et al.  Src Induces Urokinase Receptor Gene Expression and Invasion/Intravasation via Activator Protein-1/p-c-Jun in Colorectal Cancer , 2007, Molecular Cancer Research.