Regulation of microRNA -22 on glycometabolism of hematopoietic stem cell TF - 1 and its molecular mecha-nism / 中华实用儿科临床杂志

ABSTRACT

Objective To study the regulation of microRNA - 22(miR - 22)on glycometabolism of hemato-poietic stem cell TF - 1 and its molecular mechanism. Methods TF - 1 cells were cultured for 2 h under hypoxic con-ditions. 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 (Glut40. 005 ± 0. 000 vs. 0. 008 ± 0. 000;PPAR - γ0. 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 ex-erts 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.