Overexpression of miR-30a Promotes Apoptosis of Leukemia K562 Cells.
10.19746/j.cnki.issn.1009-2137.2019.02.014
- Author:
Min XU
1
;
Wen-Wan GAO
1
;
Yu-Jie LUO
1
;
Yi WANG
1
;
Kun TAO
2
Author Information
1. Department of Immunology, College of Basic Medical Science ,Chongqing Medical University, Chongqing 400016, China.
2. Department of Immunology, College of Basic Medical Science ,Chongqing Medical University, Chongqing 400016, China,E-mail: taokun68@126.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Proliferation;
Fusion Proteins, bcr-abl;
Humans;
K562 Cells;
Leukemia, Myelogenous, Chronic, BCR-ABL Positive;
MicroRNAs;
genetics
- From:
Journal of Experimental Hematology
2019;27(2):396-402
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the pro-apoptotic effect and mechanism of miR-30a overexpression on chronic myeloid leukemia K562 cells.
METHODS:The k562 cells were transfected with the recombinant plasmid pEGFP-pre-miR-30a, the real-time quantitative PCR was used to detect the level of miR-30a and BCR/ABL, and then the cell apoptosis was assessed by flow cytometry with AnnexinV-FITC/PI double staining. Western blot was used to detect the expression of BCR/ABL protein,apoptosis-related protein BCL-2 and BAX, PTEN, AKT and p-AKT.
RESULTS:Sequencing and digestion map indicated that the recombinant plasmid was constructed successfully. Compared with 2 control groups, the miR-30a expression in k562 cells transfected with recombinant plasmid pEGFP-pre-miR-30a was obviously up-regulated. The expression of BCR/ABL mRNA and BCR/ABL protein was both significantly down-regulated. Apoptotic rate was significantly enhanced (both P<0.05),and the expression of anti-apoptotic protein BCL-2 was down-regulated while the expression of pro-apoptotic protein BAX was up-regulated. The level of PTEN was significantly up-regulated in omparison with control groups,no variation was found in total AKT, but the expression of p-AKT was down-regulated.
CONCLUSION:The overexpression of miR-30a is abled to down-regulate the level of BCR/ABL mRNA and BCR/ABL protein, and increase apoptotic rate, its mechanism may be related with inhibition of the activity of BCR/ABL-PTEN/AKT signaling pathway.
