Functional microRNA is transferred between glioma cells.
MicroRNAs (miRNA) are single-stranded 17- to 27-nucleotide RNA molecules that regulate gene expression by posttranscriptional silencing of target mRNAs. Here, we transformed rat 9L gliosarcoma cells to express cel-miR-67, a miRNA that lacks homology in rat. Coculture of these cells with cells that expressed a luciferase reporter that contained a complementary sequence to cel-miR-67 resulted in significant suppression of luciferase expression. This effect was also observed in the U87-MG human glioma cell line. Moreover, luciferase suppression was inhibited by the addition of carbenoxolone to cocultures, suggesting that gap junction communication regulates intercellular transfer of miRNA. Finally, in situ hybridization revealed the presence of cel-miR-67 in cel-miR-67-null 9L cells after coculture with cel-miR-67-expressing cells. Our data show that miRNA transcribed in glioma cells can be transferred to adjacent cells and induces targeted inhibition of protein expression in the acceptor cells. These findings reveal a novel mechanism of targeted intercellular protein regulation between brain tumor cells.
Katakowski
Mark
M
Department of Neurology, Henry Ford Hospital, Detroit, Michigan 48202, USA.
Buller
Benjamin
B
Wang
Xinli
X
Rogers
Thomas
T
Chopp
Michael
M
D000818:Animals; D000897:Anti-Ulcer Agents / Q000494:pharmacology; D001932:Brain Neoplasms / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D002229:Carbenoxolone / Q000494:pharmacology; D002450:Cell Communication / Q000187:drug effects; D018920:Coculture Techniques; D017629:Gap Junctions / Q000502:physiology; D005910:Glioma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D018316:Gliosarcoma / Q000235:genetics* / Q000378:metabolism / Q000473:pathology; D049452:Green Fluorescent Proteins / Q000378:metabolism; D015151:Immunoblotting; D017403:In Situ Hybridization; D008156:Luciferases / Q000378:metabolism; D035683:MicroRNAs / Q000502:physiology*; D012333:RNA, Messenger / Q000235:genetics; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D000897:Anti-Ulcer Agents; D035683:MicroRNAs; D012333:RNA, Messenger; D049452:Green Fluorescent Proteins; D008156:Luciferases; D002229:Carbenoxolone