MicroRNA miR-328 regulates zonation morphogenesis by targeting CD44 expression.
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.
Sunnybrook Research Institute, Sunnybrook Health Sciences Centre, Toronto, Canada.
Chia-Hui
CH
Wang
Daniel Y
DY
Lee
Zhaoqun
Z
Deng
Zina
Z
Jeyapalan
Shao-Chen
SC
Lee
Shireen
S
Kahai
Wei-Yang
WY
Lu
Yaou
Y
Zhang
Burton B
BB
Yang
D000818:Animals; D015153:Blotting, Western; D002196:Capillaries / Q000254:growth & development; D002448:Cell Adhesion; D005434:Flow Cytometry; D018960:Hyaluronan Receptors / Q000378:metabolism*; D007150:Immunohistochemistry; D051379:Mice; D008819:Mice, Nude; D035683:MicroRNAs / Q000235:genetics*; D009024:Morphogenesis; D012313:RNA / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction
D018960:Hyaluronan Receptors; D035683:MicroRNAs; D012313:RNA