MicroRNA miR-328 regulates zonation morphogenesis by targeting CD44 expression

PLoS One. 2008 Jun 18;3(6):e2420. doi: 10.1371/journal.pone.0002420.


Morphogenesis is crucial to initiate physiological development and tumor invasion. Here we show that a microRNA controls zonation morphogenesis by targeting hyaluronan receptor CD44. We have developed a novel system to study microRNA functions by generating constructs expressing pre-miRNAs and mature miRNAs. Using this system, we have demonstrated that expression of miR-328 reduced cell adhesion, aggregation, and migration, and regulated formation of capillary structure. Protein analysis indicated that miR-328 repressed CD44 expression. Activities of luciferase constructs harboring the target site in CD44, but not the one containing mutation, were repressed by miR-328. Zonation morphogenesis appeared in cells transfected by miR-328: miR-328-transfected cells were present on the surface of zonating structures while the control cells stayed in the middle. MiR-328-mediated CD44 actions was validated by anti-CD44 antibody, hyaluronidase, CD44 siRNA, and CD44 expression constructs. In vivo experiments showed that CD44-silencing cells appeared as layers on the surfaces of nodules or zonating structures. Immuno-histochemistry also exhibited CD44-negative cells on the surface layers of normal rat livers and the internal zones of Portal veins. Our results demonstrate that miR-328 targets CD44, which is essential in regulating zonation morphogenesis: silencing of CD44 expression is essential in sealing the zonation structures to facilitate their extension and to inhibit complex expansion.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Blotting, Western
  • Capillaries / growth & development
  • Cell Adhesion
  • Flow Cytometry
  • Hyaluronan Receptors / metabolism*
  • Immunohistochemistry
  • Mice
  • Mice, Nude
  • MicroRNAs / genetics*
  • Morphogenesis*
  • RNA / genetics
  • Reverse Transcriptase Polymerase Chain Reaction


  • Hyaluronan Receptors
  • MicroRNAs
  • RNA