Effect of MiR-142-3p Targeting HOXA5 on Proliferation, Cycle Arrest and Apoptosis of Acute B Lymphocytic Leukemia Cells.
10.19746/j.cnki.issn.1009-2137.2021.04.011
- Author:
Wen-Cui SHEN
1
;
Yu-Wei SHI
2
Author Information
1. Department of Hematology, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830063, Xinjiang Uygur Autonomous Region, China.
2. Department of Hematology, The Second Affiliated Hospital of Xinjiang Medical University, Urumqi 830063, Xinjiang Uygur Autonomous Region, China,E-mail:yyfivedentist@163.com.
- Publication Type:Journal Article
- MeSH:
Apoptosis;
Cell Line, Tumor;
Cell Proliferation;
Genes, Homeobox;
Homeodomain Proteins/genetics*;
Humans;
Leukemia, B-Cell/genetics*;
MicroRNAs/genetics*
- From:
Journal of Experimental Hematology
2021;29(4):1085-1092
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect and molecular mechanism of miR-142-3p to the proliferation, cycle and apoptosis of acute B lymphocytic leukemia (B-ALL) cells by regulating the homeobox gene 5 (HOXA5) expression.
METHODS:Real-time fluorescence quantitative PCR was used to detect the expression levels of miR-142-3p and HOXA5 in human B-ALL cell Nalm6 cell line and human B lymphoblast Hmy2-cir cells. Nalm6 was transfected by using liposome transfection technology, miR-142-3p mimic, pcDNA-HOXA5 overexpression plasmid, miR-142-3p mimic+pcDNA-HOXA5 overexpression plasmid, and control. The binding site of HOXA5 and miR-142-3p was predicted according to microRNA.org, and the targeting relationship between miR-142-3p and HOXA5 gene was detected by double luciferase reporter gene experiment. The effect of miR-142-3p to the proliferation of Nalm6 cells was detected using the Cell Counting Box-8 (CCK-8) method and cell clone formation experiments. Flow cytometry was used to detect the effects of miR-142-3p to cell cycle distribution and apoptosis of Nalm6 cells. The expression levels of cell cycle-related proteins, including G
RESULTS:Compared with Hmy2-cir cells, miR-142-3p showed low expression in Nalm6 cells and HOXA5 showed high expression (P<0.05). MiR-142-3p and HOXA5 3'-UTR showed complementary binding regions, the luciferase activity of miR-142-3p mimic and wild-type HOXA5 3'-UTR was significantly lower than that of miR-142-3p negative control and wild-type HOXA5 3'-UTR (P<0.05). The proliferation of Nalm6 cells and the number of cell clones could be inhibited by miR-142-3p mimic after 48 and 72 hours of transfection (P<0.05), which causing G
CONCLUSION:MiR-142-3p can inhibit the proliferation of Nalm6 cells by targeting down-regulation the expression of HOXA5, arrest the G
