Effect of Silencing SET Gene on Acute Promyelocytic Leukemia NB4-R1 Cells.
- Author:
Yuan WANG
1
;
Mei ZHANG
2
;
Peng-Cheng HE
3
;
Jun QI
1
;
Yan-Feng LIU
1
;
Hua-Chao ZHU
1
Author Information
- Publication Type:Journal Article
- MeSH: Cell Cycle; Cell Line, Tumor; Gene Silencing; Genetic Vectors; HEK293 Cells; Histone Chaperones; genetics; Humans; Lentivirus; Leukemia, Promyelocytic, Acute; genetics; pathology; Protein Phosphatase 2; metabolism; RNA, Messenger; RNA, Small Interfering; Transcription Factors; genetics; Transfection
- From: Journal of Experimental Hematology 2016;24(1):41-45
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of silencing SET gene on the biological characteristics of acute promyelocytic leukemia NB4-R1 cells.
METHODSThe 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.
RESULTSThe 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.
CONCLUSIONThe 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.
