Regulation of microRNA-22 on glycometabolism of hematopoietic stem cell TF-1 and its molecular mechanism

Abstract

Objective To study the regulation of microRNA-22(miR-22) on glycometabolism of hematopoietic stem cell TF-1 and its molecular mechanism. Methods TF-1 cells were cultured for 2 h under hypoxic conditions.The expression levels of Glut4 and miR-22 was detected by quantitative real-time PCR(qRT-PCR). The sgRNA vector of the miR-22 gene was constructed and miR-22 gene of TK-1 cells was knocked out by the CRISPR/Cas9 technique.Overexpression vectors were constructed, and miR-22 knocked-out cells were introduced to overexpress miR-22, the expression of miR-22 was detected by qRT-PCR and the expression levels of Glut4 and PPAR-γ were detected by qRT-PCR and Western blot. Results Compared with the control group, the expression of miR-22 in TF-1 cells decreased (0.015±0.000 vs. 0.056±0.001) and the expression of Glut4 (0.351±0.038 vs. 0.152±0.005) and PPAR-γ (0.421±0.017 vs. 0.248±0.008) increased, when TF-1 cells were cultured for 2 h under hypoxic conditions, and the differences were statistically significant (all P<0.05). Compared with the control group, the expression levels of Glut4 (0.019±0.00 vs. 0.008±0.000) and PPAR-γ (0.038±0.001 vs. 0.019±0.000) were significantly increased after miR-22 gene silencing, and they were significantly decreased (Glut4: 0.005±0.000 vs. 0.008±0.000; PPAR-γ: 0.137±0.000 vs. 0.019±0.000) after overexpression of miR-22, and the differences were statistically significant (all P<0.05). Conclusions It suggests that miR-22 exerts a negative regulation on glycometabolism of hematopoietic stem cell TF-1 by downregulating the expression of PPAR-γ.A new regulatory factor of TF-1 glycometabolism and the mechanisms are identified, which has provided new ideas for the targeted medication of diseases induced by hematopoietic stem cell dysfunction. Key words: Hematopoietic stem cells; Glycometabolism; MicroRNA; Peroxisome proliferators-activated receptor-γ