Effect of MiRNA-145 on Apoptosis of Leukemia HuT 78 Cells and Its Mechanism.
10.7534/j.issn.1009-2137.2020.01.018
- Author:
Zhi-Hui LI
1
;
Yan-Ping ZHANG
1
;
Peng-Tao XING
1
;
Xin-Rong ZHAN
2
Author Information
1. Department of Hematology, Xinxiang Central Hospital, Xinxiang 453000, Henan Province, China.
2. Department of Hematology, Xinxiang Central Hospital, Xinxiang 453000, Henan Province, China,E-mail: zhanxinrong3137161@163.com.
- Publication Type:Journal Article
- From:
Journal of Experimental Hematology
2020;28(1):104-109
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect and mechanism of miRNA-145 on leukemic cell apoptosis.
METHODS:After transfection of miRNA-145 mimic and negative control mimic in leukemia cells by Lipofectamine 2000 liposome, the MTT assay was used to detect the effect of miRNA-145 on cell proliferation. Flow cytometry was used to detect the effect of miRNA-145 on cell cycle and apoptosis. Western blotting assay was used to detect the expression levels of BCL-2, CDK6, Cyclin D1, BAX, PI3K p-PI3K, p-AKT and AKT.
RESULTS:The relative level of microRNA in HuT 78 cells transfected with miRNA-145 was 2.3±02, which was significantly higher than that in blank control group and miRNA-NC group (P＜0.05). MTT assay showed that the proliferation level of HuT 78 cells transfected with miRNA-145 mimic was significantly lower than that of blank control and miRNA-NC group (P＜0.05). Flow cytometry showed that the cells at G/G, S and G2/M phase of HuT 78 cells were significantly decreased after transfection with miRNA-145 mimic (P＜0.05). Annexin V/PI double staining assay showed that the apoptosis rate of HuT 78 cells was 17.6%±3.4%，which was significantly higher than that in blank control group and miRNA-NC group (P＜0.05). Western blot showed that the expression levels of BCL-2, CDK6 and Cyclin D1 in HuT 78 cells were significantly lower than those in blank control and miRNA-NC group (P＜0.05), and BAX expression in HuT 78 cells was significantly higher than that in blank control and miRNA-NC group (P＜0.05). Western blot showed that expression of PI3K, p-PI3K, AKT and p-AKT in HuT 78 cells transfected with miRNA-145 mimic were significantly lower than that in blank control and miRNA-NC group (P＜0.05).
CONCLUSION:Upregulation of miRNA-145 may inhibit the proliferation of leukemia cells and promote the apoptosis, which may be related with the inhibition of PI3K/AKT signaling pathway.
