Expression of CD44 3′-untranslated region regulates endogenous microRNA functions in tumorigenesis and angiogenesis

The non-coding 3′-untranslated region (UTR) plays an important role in the regulation of microRNA (miRNA) functions, since it can bind and inactivate multiple miRNAs. Here, we show the 3′-UTR of CD44 is able to antagonize cytoplasmic miRNAs, and result in the increased translation of CD44 and downstream target mRNA, CDC42. A series of cell function assays in the human breast cancer cell line, MT-1, have shown that the CD44 3′-UTR inhibits proliferation, colony formation and tumor growth. Furthermore, it modulated endothelial cell activities, favored angiogenesis, induced tumor cell apoptosis and increased sensitivity to Docetaxel. These results are due to the interaction of the CD44 3′-UTR with multiple miRNAs. Computational algorithms have predicted three miRNAs, miR-216a, miR-330 and miR-608, can bind to both the CD44 and CDC42 3′-UTRs. This was confirmed with luciferase assays, western blotting and immunohistochemical staining and correlated with a series of siRNA assays. Thus, the non-coding CD44 3′-UTR serves as a competitor for miRNA binding and subsequently inactivates miRNA functions, by freeing the target mRNAs from being repressed.

[1]  Burton B. Yang,et al.  A 3′-Untranslated Region (3′UTR) Induces Organ Adhesion by Regulating miR-199a* Functions , 2009, PloS one.

[2]  Gregory J. Hannon,et al.  MicroRNA-dependent localization of targeted mRNAs to mammalian P-bodies , 2005, Nature Cell Biology.

[3]  L. Zon,et al.  Cdc42Hs, but Not Rac1, Inhibits Serum-stimulated Cell Cycle Progression at G1/S through a Mechanism Requiring p38/RK* , 1997, The Journal of Biological Chemistry.

[4]  E. Ruoslahti,et al.  The C-type lectin domains of lecticans, a family of aggregating chondroitin sulfate proteoglycans, bind tenascin-R by protein-protein interactions independent of carbohydrate moiety. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[5]  Burton B. Yang,et al.  The ability of versican to simultaneously cause apoptotic resistance and sensitivity. , 2007, Cancer research.

[6]  R. Stallings,et al.  Differential patterns of microRNA expression in neuroblastoma are correlated with prognosis, differentiation, and apoptosis. , 2007, Cancer research.

[7]  Vasudevan Seshadri,et al.  Translational control by the 3'-UTR: the ends specify the means. , 2003, Trends in biochemical sciences.

[8]  C. Wollheim,et al.  MicroRNAs: 'ribo-regulators' of glucose homeostasis , 2006, Nature Medicine.

[9]  Yaou Zhang,et al.  MicroRNA MiR-17 retards tissue growth and represses fibronectin expression , 2009, Nature Cell Biology.

[10]  M. Glimcher,et al.  Receptor-Ligand Interaction Between CD44 and Osteopontin (Eta-1) , 1996, Science.

[11]  C. Croce,et al.  MicroRNA gene expression deregulation in human breast cancer. , 2005, Cancer research.

[12]  M. Akens,et al.  The effect of versican G3 domain on local breast cancer invasiveness and bony metastasis , 2007, Breast Cancer Research.

[13]  M. Hentze,et al.  Molecular mechanisms of translational control , 2004, Nature Reviews Molecular Cell Biology.

[14]  V. Kosma,et al.  Versican in epithelial ovarian cancer: Relation to hyaluronan, clinicopathologic factors and prognosis , 2003, International journal of cancer.

[15]  Oliver Hobert,et al.  A microRNA controlling left/right neuronal asymmetry in Caenorhabditis elegans , 2003, Nature.

[16]  T. Dalmay,et al.  MicroRNAs and the hallmarks of cancer , 2006, Oncogene.

[17]  H. Hosick,et al.  Blocking HER-2/HER-3 function with a dominant negative form of HER-3 in cells stimulated by heregulin and in breast cancer cells with HER-2 gene amplification. , 2000, Cell growth & differentiation : the molecular biology journal of the American Association for Cancer Research.

[18]  H. Ruohola-Baker,et al.  Stem cell division is regulated by the microRNA pathway , 2005, Nature.

[19]  E. Cuppen,et al.  Limitations and possibilities of small RNA digital gene expression profiling , 2009, Nature Methods.

[20]  E. Bartnik,et al.  A Requirement for the CD44 Cytoplasmic Domain for Hyaluronan Binding, Pericellular Matrix Assembly, and Receptor-mediated Endocytosis in COS-7 Cells* , 2002, The Journal of Biological Chemistry.

[21]  T. Taher,et al.  Cross-talk between CD44 and c-Met in B cells. , 1999, Current topics in microbiology and immunology.

[22]  Christophe Lemetre,et al.  MicroRNA signatures predict oestrogen receptor, progesterone receptor and HER2/neu receptor status in breast cancer , 2009, Breast Cancer Research.

[23]  Shuhong Zhao,et al.  Discovery of Porcine microRNAs and Profiling from Skeletal Muscle Tissues during Development , 2008, PloS one.

[24]  O. Gotoh,et al.  Species-specific variation of alternative splicing and transcriptional initiation in six eukaryotes. , 2005, Gene.

[25]  R. Shiekhattar,et al.  TRBP recruits the Dicer complex to Ago2 for microRNA processing and gene silencing , 2005, Nature.

[26]  Yaou Zhang,et al.  MicroRNA miR-328 Regulates Zonation Morphogenesis by Targeting CD44 Expression , 2008, PloS one.

[27]  E. Danen,et al.  Expression of cd44 splice variants in human cutaneous melanoma and melanoma cell lines is related to tumor progression and metastatic potential , 1995, International journal of cancer.

[28]  Jie Yang,et al.  MicroRNA miR-93 promotes tumor growth and angiogenesis by targeting integrin-β8 , 2011, Oncogene.

[29]  I. Stamenkovic,et al.  CD44 is the principal cell surface receptor for hyaluronate , 1990, Cell.

[30]  V. Narry Kim,et al.  Functional links between clustered microRNAs: suppression of cell-cycle inhibitors by microRNA clusters in gastric cancer , 2009, Nucleic acids research.

[31]  G. Ruvkun.  Clarifications on miRNA and Cancer , 2006, Science.

[32]  C. Klinge,et al.  Estradiol downregulates miR-21 expression and increases miR-21 target gene expression in MCF-7 breast cancer cells , 2009, Nucleic acids research.

[33]  P. Isaacson,et al.  Immunoproliferative small-intestinal disease. An immunohistochemical study. , 1989, The American journal of surgical pathology.

[34]  Jonathan Sleeman,et al.  CD44 Acts Both as a Growth‐ and Invasiveness‐Promoting Molecule and as a Tumor‐Suppressing Cofactor , 2000, Annals of the New York Academy of Sciences.

[35]  U. Günthert,et al.  Expression of cell adhesion molecules alpha-2, alpha-5 and alpha-6 integrin, E-cadherin, N-CAM and CD-44 in renal cell carcinomas , 1993, Virchows Archiv A.

[36]  Burton B. Yang,et al.  MicroRNA-378 promotes cell survival, tumor growth, and angiogenesis by targeting SuFu and Fus-1 expression , 2007, Proceedings of the National Academy of Sciences.

[37]  K. Pienta,et al.  Establishment of immortalized Copenhagen rat prostate endothelial cell lines. , 1995, In vivo.

[38]  C. Burge,et al.  Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets , 2005, Cell.

[39]  S. Fox,et al.  Normal human tissues, in addition to some tumors, express multiple different CD44 isoforms. , 1994, Cancer research.

[40]  John W M Martens,et al.  Four miRNAs associated with aggressiveness of lymph node-negative, estrogen receptor-positive human breast cancer , 2008, Proceedings of the National Academy of Sciences.