Large numbers of microRNAs (miRNAs) have been found to be dysregulated in β-thalassemia (β-thal), but their roles in β-thal are poorly reported. The purpose of this study was to investigate the clinical significance of miR-17-3p in β-thal, and to elucidate its regulatory effect on K562 cell proliferation and erythroid differentiation as well as γ-globin expression.17 patients with β-thal intermedia and major and 17 healthy controls were selected and peripheral blood samples were collected from the participants. The expression of miR-17-3p and B cell lymphoma 11A (BCL11A) were detected by qRT-PCR and Western blotting. The regulation of miR-17-3p on BCL11A was evaluated by luciferase reporter gene assay. Cell proliferation, cell cycle, cell apoptosis and erythroid differentiation was assessed CCK-8 and Flow cytometry.The data demonstrated that upregulated miR-17-3p expression is associated with patients with β-thal intermedia and major. Although miR-17-3p has no significant effect on cell proliferation and erythroid differentiation, miR-17-3p promotes γ-globin expression by targeting BCL11A, suggesting that miR-17-3p may be an promising miRNA for the treatment of β-thal.