MiR-223-3p regulates erythropoiesis by targeting TGFBR3/Smad signaling pathway in hemoglobin H-Constant Spring disease

• Published in: Annals of medicine (Helsinki) Vol. 57; no. 1; p. 2530690
• Main Authors: Lan, Lan, Wang, Xinyu, Zhang, Simeng, Zeng, Dan, Mo, Limin, Feng, Qiao, Li, Jun, Zhu, Chunjiang
• Format: Journal Article
• Language: English
• Published: England Taylor & Francis 01.12.2025; Taylor & Francis Group
• Subjects: Adult; Apoptosis - genetics; Case-Control Studies; Cell Proliferation; Down-Regulation; Erythropoiesis - genetics; Female; HbH-CS disease; Hematology; Humans; Male; MicroRNAs - genetics; MicroRNAs - metabolism; Middle Aged; miR-223-3p; Proteoglycans; Receptors, Transforming Growth Factor beta - genetics; Receptors, Transforming Growth Factor beta - metabolism; Signal Transduction - genetics; Smad2 Protein - genetics; Smad2 Protein - metabolism; Smad3 Protein - genetics; Smad3 Protein - metabolism; TGFBR3; Thalassemia; TGFBR3; Thalassemia; HbH-CS disease; miR-223-3p
• DOI: 10.1080/07853890.2025.2530690
• ISSN: 0785-3890; 1365-2060

MicroRNAs (miRNAs) have emerged as regulators of pathogenesis in hematopoiesis by targeting post-transcription mRNA regulation. However, the involvement of miRNAs in hemoglobin H-Constant Spring (HbH-CS) disease remains unclear. The present study analyzed miRNAs differential expression profile between patients with HbH-CS disease and healthy subjects by Arraystar Human small RNA Microarray. The differential expression profiles of miRNAs between HbH-CS patients and healthy individuals were analyzed using Arraystar human small RNA microarrays. Quantitative real-time PCR and Western blot were used to detect the mRNA and protein expression. Bioinformatics methods were used to predict the target genes of miRNAs, and the regulatory relationship between miRNAs and target genes was verified by dual-luciferase reporter assay. Cell apoptosis was detected by flow cytometry, and cell proliferative capacity was detected by CC K-8 assay. MiR-223-3p was significantly down-regulated in HbH-CS patients. Dual-luciferase reporter assay demonstrated that miR-223-3p specifically targeted TGFBR3. The experiments showed that miR-223-3p inversely regulated the expression levels of TGFBR3, Smad2/3, and P-Smad2. We found that overexpression of miR-223-3p in K562 cells significantly promoted cellular proliferation and inhibited cellular apoptosis by suppressing expression of TGFBR3, Smad2/3, and P-Smad2. Moreover, up-regulation of miR-223-3p in induced K562 cells increased the expression levels of hemoglobin and mean corpuscular hemoglobin. In conclusion, this study indicated that the interaction between miR-223-3p and TGFBR3/Smad signaling pathway might affect erythropoiesis in HbH-CS disease, which can provide more information for the treatment of patients.