The Relationship between PPP2R5C and Molt-4 Cell Viability, HSP90-GR Signal in Childhood Acute T Lymphocytic Leukemia.
10.19746/j.cnki.issn.1009-2137.2022.01.014
- Author:
Lei LIU
1
;
Hai-Tao LI
1
;
Hua-Yue ZHENG
1
;
Hui-Bing DANG
2
Author Information
1. The Second Department of Pediatrics, The First Affiliated Hospital of Nanyang Medical College, Nanyang 473000, Henan Province, China.
2. Department of Hematology, The First Affiliated Hospital of Nanyang Medical College, Nanyang 473000, Henan Province, China,E-mail: bluewine_ll@sina.com.
- Publication Type:Journal Article
- Keywords:
PPP2R5C;
cell viability;
childhood acute T lymphocytic leukemia;
glucocorticoid receptor;
heat shock protein 90
- MeSH:
Apoptosis;
Cell Line, Tumor;
Cell Proliferation;
Cell Survival;
Child;
HSP90 Heat-Shock Proteins;
Humans;
Precursor T-Cell Lymphoblastic Leukemia-Lymphoma;
RNA, Small Interfering;
Receptors, Glucocorticoid
- From:
Journal of Experimental Hematology
2022;30(1):84-91
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:To investigate the effect of PPP2R5C to the activity of Molt-4 cells in childhood acute T lymphocytic leukemia and its mechanism.
METHODS:The small interfering RNA (siRNA) technology targeting PPP2R5C gene was used to down-regulate the expression of PPP2R5C in Molt-4 cells. At the same time, a blank control group, a negative control group and a 17-DMAG group were set up. The cells in the negative control group were transfected with siRNA-NC, the cells in 17-DMAG group were treated with the HSP90 inhibitor 17-DMAG at a final concentration of 6.4 μmol/L for 48 h. Real-time fluorescent quantitative PCR (RT-qPCR) and Western blot were used to detect transfection efficiency; CCK-8 method was used to detect the proliferation activity of the cells in each group, EdU was used to detect the proliferation level of the cells in each group, flow cytometry was used to detect the cell cycle distribution ratio of the cells in each group, Annexin V-FITC/PI staining was used to detect the apoptosis of the cell, RT-qPCR and Western blot were used to detect the expression changes of heat shock protein 90 (HSP90) and glucocorticoid receptor (GR) of the cells in each group.
RESULTS:After Molt-4 cells were transfected with siRNA-PPP2R5C, the expression of PPP2R5C mRNA and protein in the cells were down-regulated significantly compared with those in the blank control group and the si-NC group (P<0.05); compared with cells in the blank control group and the si-NC group, the proliferation activity of the cells in the siRNA-PPP2R5C group and the 17-DMAG group significantly decreased (P<0.05), and the rate of EdU positive cells was significantly reduced (P<0.05); the proportion of the cells in G1 phase decreased while the proportion of the cells in G2 phase increased (P<0.05), the apoptosis rate of the cells also increased significantly (P<0.05); in addition, the expression of PPP2R5C mRNA and protein of the cells in siRNA-PPP2R5C group was significantly down-regulated compared with those in the blank control group and si-NC group (P<0.05). The expressions of PPP2R5C mRNA and protein in the 17-DMAG group were also significantly down-regulated compared with those in the blank control group and si-NC group (P<0.05).
CONCLUSION:Down-regulation of PPP2R5C gene expression can inhibit Molt-4 cell activity in childhood acute T lymphocytic leukemia, block the cells in G2 phase, and promote cell apoptosis, the mechanism may be related to the inhibition of HSP90-GR signaling pathway.
