Chronic Myeloid Leukaemia (CML) is a
myeloproliferative disorder characterized by the expression of the
oncoprotein, Bcr-Abl
kinase. CCN3 normally functions as a negative growth regulator, but it is downregulated in CML, the mechanism of which is not known.
MicroRNAs (
miRNAs) are small non-coding RNAs, which negatively regulate protein translation by binding to the complimentary sequences of the
3' UTR of messenger RNAs. Deregulated
miRNA expression has emerged as a hallmark of
cancer. In CML, BCR-ABL upregulates oncogenic
miRNAs and downregulates tumour suppressor
miRNAs favouring leukaemic transformation. We report here that the downregulation of CCN3 in CML is mediated by BCR-ABL dependent
miRNAs. Using the CML cell line K562, we profiled
miRNAs, which are BCR-ABL dependent by transfecting K562 cells with anti-BCR-ABL
siRNA.
MiRNA expression levels were quantified using the Taqman Low Density
miRNA array platform. From the
miRNA target prediction databases we identified
miRNAs that could potentially bind to CCN3
mRNA and reduce expression. Of these, miR-130a, miR-130b, miR-148a, miR-212 and miR-425-5p were significantly reduced on BCR-ABL knockdown, with both miR-130a and miR-130b decreasing the most within 24 h of
siRNA treatment. Transfection of mature sequences of miR-130a and miR-130b individually into BCR-ABL negative HL60 cells resulted in a decrease of both CCN3
mRNA and
protein. The reduction in CCN3 was greatest with overexpression of miR-130a whereas miR-130b overexpression resulted only in marginal repression of CCN3. This study shows that
miRNAs modulate CCN3 expression. Deregulated
miRNA expression initiated by BCR-ABL may be one mechanism of downregulating CCN3 whereby leukaemic cells evade negative growth regulation.