Antiproliferative effect of silencing LSD1 gene on Jurkat cell line and its mechanism.
- Author:
Shiwei HAN
1
;
Yiqun HUANG
;
Ruiji ZHENG
Author Information
- Publication Type:Journal Article
- MeSH: Acetylation; Apoptosis; Caspase 3; metabolism; Cell Cycle; Cell Line, Tumor; Cell Proliferation; Down-Regulation; Histone Demethylases; genetics; Histones; metabolism; Humans; Jurkat Cells; Methylation; Proto-Oncogene Proteins c-bcl-2; metabolism; RNA Interference; RNA, Messenger; RNA, Small Interfering; Transfection
- From: Chinese Journal of Hematology 2016;37(1):56-60
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo investigate the effect of silencing LSD1 gene by RNA interference on the proliferation, apoptosis on human lymphocytic leukemia Jurkat cell line and its mechanism.
METHODSThe hairpin- like oligonucleotide sequences targeting LSD1 gene was transfected into Jurkat cells by lipofectamine(TM) 2000. The LSD1 mRNA and protein were detected by RQ- PCR and Western blot. Cell growth was determined by MTT. Cell apoptosis was analyzed by flow cytometry. The expression of Bcl-2, Bax, procaspase- 3, and histone H3K4me, H3K4me2, H3K4me3, Act- H3, H3K9me were detected by Western blot.
RESULTSLSD1 mRNA was markedly suppressed by the shRNA targeting LSD1. LSD1 shRNA suppressed the proliferation and induced cells apoptosis of Jurkat cells. The cell apoptotic rate was (41.34±3.58)%, (3.45±1.54)%, (1.76±0.52)% in LSD1 shRNA, Neg-shRNA and Blank respectively, the difference among them was statistically significant (P<0.05). LSD1 shRNA down- regulated the expressions of Bcl- 2 and procaspase- 3, and up- regulated the expression of Bax. The methylation of H3K4me1, me2 and acetylation of Act- H3 improved without change of the methylation of H3K4me3.
CONCLUSIONSDeplete of LSD1 gene maybe through modifying the methylation of histone H3K4 to promote the cell apoptosis and inhibit cell growth in Jurkat cell line.
