- Author:
Jin-Xia CAO
1
;
Feng WEN
1
;
Hao WANG
2
;
Pei LIAO
1
;
Jun-Jun LI
3
Author Information
- Publication Type:Journal Article
- MeSH: CCAAT-Enhancer-Binding Protein-alpha; Cell Proliferation; Cell Survival; Humans; K562 Cells; Leukemia, Myelogenous, Chronic, BCR-ABL Positive; MicroRNAs; Transfection; Up-Regulation
- From: Journal of Experimental Hematology 2015;23(2):350-355
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEOur research was aim to investigate the effect of microRNA-328 (miR-328) on proliferation of chronic myeloid leukemia(CML) cell K562 and the mediated effect of C/EBPα.
METHODSThe eukaryotic expression vectors of miR-328 targeting gene and suppressor gene (hsa-miR-328 and hsa-miR-328-inhibitor) were constructed, and transfected into K562 cells respectively. The mRNA expression levels of miR-328 and C/EBP α were detected by real-time fluorescence quantitative RT-PCR; C/EBP α protein expression was detected by Western blot; CCK-8 was used to estimate the cell viability.
RESULTSThe recombinant genes of hsa-miR-328 and hsa-miR-328-inhibitor were successfully constructed and transfected into K562 cells. Fluorescent cells were observed after 24 h, and the visible fluorescence cells were gradually increased after 48 h or 72 h, the miR-328 showed no effect on the mRNA expression of C/EBPα detected by RT-PCR. Meanwhile, miR-328 showed recovering effect on C/EBPα translation and inhibition of K562 cells proliferation.
CONCLUSIONmiR-328 has been successfully constructed and transfected into K562 cells, miR-328 inhibits the proliferation of K562 cells by up-regulation of C/EBPα.