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[The Relationship between PPP2R5C and Molt-4 Cell Viability, HSP90-GR Signal in Childhood Acute T Lymphocytic
Lei Liu1, Hai-Tao Li1, Hua-Yue Zheng1
1The Second Department of Pediatrics, The First Affiliated Hospital of Nanyang Medical College, Nanyang 473000, Henan Province, China.
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.
Insights
Down-regulating PPP2R5C inhibits Molt-4 cell activity in childhood acute T lymphocytic leukemia by promoting apoptosis and cell cycle arrest. This mechanism may involve the HSP90-GR signaling pathway.
Area of Science:
- Oncology
- Molecular Biology
- Cell Biology
Background:
- Childhood acute T lymphocytic leukemia (T-ALL) is an aggressive cancer.
- Understanding the molecular mechanisms driving T-ALL progression is crucial for developing effective therapies.
- PPP2R5C has been implicated in cellular processes, but its role in T-ALL is not fully understood.
Purpose of the Study:
- To investigate the effect of PPP2R5C on Molt-4 cells in childhood T-ALL.
- To elucidate the underlying mechanism of PPP2R5C's action in T-ALL.
- To explore potential therapeutic targets by modulating PPP2R5C expression.
Main Methods:
- Small interfering RNA (siRNA) was used to down-regulate PPP2R5C expression in Molt-4 cells.
- Cell proliferation was assessed using CCK-8 and EdU assays.
- Cell cycle distribution and apoptosis were analyzed by flow cytometry (Annexin V-FITC/PI staining).
- Expression of PPP2R5C, HSP90, and GR was quantified using RT-qPCR and Western blot.
Main Results:
- Down-regulation of PPP2R5C significantly reduced Molt-4 cell proliferation and EdU-positive rates.
- PPP2R5C inhibition led to G2 phase cell cycle arrest and increased apoptosis.
- The expression of HSP90 and GR was altered following PPP2R5C down-regulation.
Conclusions:
- Down-regulation of PPP2R5C inhibits Molt-4 cell activity in T-ALL.
- PPP2R5C inhibition induces G2 cell cycle arrest and promotes apoptosis.
- The mechanism may involve the inhibition of the HSP90-GR signaling pathway.
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