Regulatory Effect of MiR-543 on Proliferation and Apoptosis of Human Leukemia Cell Line K562 and Its Mechanism.
10.19746/j.cnki.issn.1009-2137.2019.03.020
- Author:
Ling SUN
1
;
Jun LU
1
;
Tan-Tan LI
1
;
Li-Jie MEN
1
;
Chuan-Cui LI
2
Author Information
1. Department of Hematology, Laiwu People's Hospital, Laiwu City 271100, Shandong Province, China.
2. Department of Hematology, Laiwu People's Hospital, Laiwu City 271100, Shandong Province, China,E-mail: 120165870@qq.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Line, Tumor;
Cell Proliferation;
Humans;
K562 Cells;
Leukemia;
MicroRNAs;
genetics
- From:
Journal of Experimental Hematology
2019;27(3):763-768
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the regulatory effect of miR-543 on proliferation and apoptosis of human leukemia cell line K562 and its mechanism.
METHODS:46 CML patients treated in our hospital from 2018.2-2018.9 was selected and enrolled in CML group, and 30 healthy persons were selected and enrolled in control group at the same time. After Lipofectamine 2000 liposome was used to transfer the miR-543 mimic and negative control (Scramble mimic) to K562 cells, CCK8 assay was used to detect the effect of miR-543 on proliferation of K562 cells; Luciferase reporter assay was used to report the targeting relationship of miR-543 to Wnt gene. Flow cytometry was used to detect the effect of miR-543 on apoptosis of K562 cells; Western blot method was used to detect the Wnt, β-catenin, BCL-2, c-MYC and BAX expression level.
RESULTS:The level of miRNA-543 in CML patients was significantly lower than that in healthy controls (P＜0.05). The expression level of miR-543 in mimic group was significantly higher than that in blank control group (P＜0.05). The Wnt gene expression level in mimic group was significantly lower than that in blank control group (P＜0.05). The expression of luciferase of Wnt wild plasmid was decreased by miR-543 mimic (P＜0.05), and the luciferase activity of Wnt mutant plasmid was not inhibited by miR-543 mimic after mutation of binding site (P＜0.05). There was no significant difference in the proliferation ability of K562 cells between the blank control group and the negative control (Scramble mimic) group (P＞0.05). The proliferation level of K562 cells in mimic group was significantly lower than that in blank control group and negative control (Scramble mimic) group (P＜0.05). Apoptotic level of K562 cells in miR-543 mimic group was significantly higher than that in blank control group and negative control (Scramble mimic) group (P＜0.05). Western blot assay showed that Wnt, β-catenin, BCL-2 and c-MYC protein levels in miR-543 mimic group were significantly lower than those in blank control group and negative control (Scramble mimic) group (P＜0.05); BAX protein level in miR-543 mimic group was higher than that in blank control group and negative control (Scramble mimic) group (P＜0.05).
CONCLUSION:miR-543 can target Wnt protein to inhibit the activity of Wnt signaling pathway, thereby inhibiting the proliferation of leukemia cells and increasing the level of apoptosis.
