Abstract: ObjectiveTo explore the role of aquaporin-1(AQP1) gene in erythroid differentiation of erythroleukemia K562cells
induced by retinoic acid (RA). MethodsK562cells were cultured in the presence of1μmol/L RA for varying lengths of time,
andγ-globin mRNA expression and hemoglobin content in the cells were detected by real-time PCR (RT-PCR) and ultraviolet
spectrophotometry, respectively, to evaluate the erythroid differentiation of K562cells. RT-PCR and Western blotting were
used to examine AQP1expression in the cells following RA treatment. A retroviral expression vector of AQP1small interfering
RNA (pSUPER-retro-puro-shAQP1) was constructed and transfected into K562cells to establish a K562cell line with stable
AQP1down-regulation (K562-shAQP1), in which the changes in γ-globin and hemoglobin expressions after RA treatment
were detected.ResultsRA treatment significantly increasedγ-globin and hemoglobin expressions in K562cells (P<0.01),
causing also significantly enhanced AQP1 mRNA and protein expressions over time (P<0.01). Transfection with the
recombinant plasmids pSuper-retro-puro-shAQP1resulted in stable AQP1suppression in K562cells (P<0.01), which showed
markedly reducedγ-globin and hemoglobin expressions after RA induction as compared to the control K562cells (P<0.01).
ConclusionsK562 cells show a significant increase of AQP1 expression after RA-induced erythroid differentiation, and
suppression of AQP1expression can partially block the effect of RA, suggesting the important role of AQP1in RA-induced
erythroid differentiation of K562cells.