Identification of uPAR Variants Acting as ceRNAs in Leukaemia Cells

SIMPLE SUMMARY: The urokinase (uPA) receptor (uPAR) concentrates proteolytic activities on the cell surface and is an adhesion receptor for vitronectin. Urokinase/Vitronectin binding to uPAR activates intracellular signals promoting cell adhesion, migration, proliferation and survival. Thus, uPAR can sustain most activities of malignant cells and, accordingly, increased uPAR expression is associated with poor prognosis in several malignancies. We previously demonstrated that, in leukaemia cells, the uPAR 3′untranslated region (3′UTR) up-regulates the expression of pro-tumoral factors by recruiting microRNAs targeting their mRNAs, thus acting as competitive endogenous RNA (ceRNA). Here, we identify 3′UTR-containing variants of uPAR mRNA in leukaemia cells and demonstrate that the over-expression of uPAR Δ5-variant mRNA promotes expression of pro-tumoral factors and increase in biological activities, probably by its ceRNA activity. On this basis, we propose that uPAR may play a crucial role in cancer biology also at mRNA level, through the ceRNA activity of its variants. ABSTRACT: The 3′untranslated region (3′UTR) of the urokinase (uPA) receptor (uPAR) mRNA can act as a competitive endogenous RNA (ceRNA) in acute myeloid leukaemia (AML) cells, promoting the expression of pro-tumoral targets, including uPAR. Here, we identified three variants of uPAR mRNA containing the 3′UTR, in KG1 and U937 leukaemia cells expressing low and high uPAR levels, respectively. Identified variants lack exon 5 (uPAR Δ5) or exon 6 (uPAR Δ6) or part of exon 6, exon 7 and part of 3′UTR (uPAR Δ6/7). uPAR Δ5 and uPAR Δ6 transcript levels were higher in U937 cells compared to KG1 cells. Both uPAR variants were expressed also in AML blasts, at higher levels as compared to CD34 hematopoietic cells from healthy donors. The presence of the 3′UTR conferred high instability to the uPAR Δ5 variant transcript, preventing its translation in protein. Overexpression of the uPAR Δ5-3′UTR variant regulated the expression of some pro-tumoral factors previously reported to be regulated by the 3′UTR of uPAR and increased KG1 cell adhesion, migration and proliferation. These results demonstrate the expression of uPAR mRNA variants containing the 3′UTR in AML cells and the ceRNA activity and the biological effects of the uPAR Δ5-3′UTR variant.