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[Effect of Silencing SET Gene on Acute Promyelocytic Leukemia NB4-R1 Cells]
Yuan Wang1, Mei Zhang1, Peng-Cheng He1
1Department of Hematology, The First Affiliated Hospital, Xi'an Jiaotong University Medical College, Xi'an 710061, Shaanxi Province, China.
Objective:
To investigate the effect of silencing SET gene on the biological characteristics of acute promyelocytic leukemia NB4-R1 cells.
Methods:
The expression vector of pGCSIL containing SET-shRNA were transfected into 293T cells by using other packaging plasmids. The supernatant of the 293T cells was harvested for lentivirus. The SET-shRNA lentiviral vector was transfected into acute promyelocytic leukemia NB4-R1 cells and a stably transfected cell line was established. Real-time quantitative PCR and Western blot were used to assay the silencing efficiency on SET gene and the expression of PP2A. The cell cycle distribution was tested by flow cytometry.
Results:
The expression of SET in experimental group statistically decreased as compared with that of the control group. The expression of PP2A was obviously raised at the level of mRNA and protein. The percentage of NB4-R1 cells in G0/G1 phase significantly increased, while the percentage of cells in S phase significantly decreased.
Conclusion:
The silencing gene in acute promyelocytic leukemia NB4-R1 cells using SET-shRNA lentiviral vector can increase the expression of PP2A and interfere of the cell cycle in NB4-R1 cells. This study has laid a experimental base for targed therapy of patients with acute promyelocytic leukemia.
Insights
Silencing the SET gene in acute promyelocytic leukemia cells increased PP2A expression and altered cell cycle progression. This provides a potential therapeutic strategy for acute promyelocytic leukemia.
Area of Science:
- Molecular Biology
- Cell Biology
- Hematology
Background:
- Acute promyelocytic leukemia (APL) is a distinct subtype of acute myeloid leukemia.
- The SET gene plays a role in the pathogenesis of APL.
- Understanding the molecular mechanisms underlying APL is crucial for developing targeted therapies.
Purpose of the Study:
- To investigate the impact of SET gene silencing on the biological characteristics of acute promyelocytic leukemia NB4-R1 cells.
- To evaluate the effect of SET gene silencing on PP2A expression and cell cycle distribution.
Main Methods:
- Lentiviral vectors containing SET-shRNA were used to establish a stably transfected NB4-R1 cell line.
- Real-time quantitative PCR and Western blot assays were employed to assess SET gene silencing and PP2A expression.
- Flow cytometry was utilized to analyze cell cycle distribution.
Main Results:
- SET gene expression was significantly decreased in the experimental group compared to the control.
- PP2A expression, at both mRNA and protein levels, was markedly increased.
- A significant increase in the percentage of NB4-R1 cells in the G0/G1 phase and a significant decrease in the S phase were observed.
Conclusions:
- SET gene silencing in NB4-R1 cells effectively increases PP2A expression and disrupts cell cycle progression.
- This study provides a foundational experimental basis for targeted therapy in acute promyelocytic leukemia patients.
- Targeting the SET gene represents a promising therapeutic avenue for APL.
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