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[Antiproliferative effect of silencing LSD1 gene on Jurkat cell line and its mechanism]
Shiwei Han1, Yiqun Huang, Ruiji Zheng
1Department of Hematology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou 363000, China.
Objective:
To investigate the effect of silencing LSD1 gene by RNA interference on the proliferation, apoptosis on human lymphocytic leukemia Jurkat cell line and its mechanism.
Methods:
The 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.
Results:
LSD1 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.
Conclusions:
Deplete of LSD1 gene maybe through modifying the methylation of histone H3K4 to promote the cell apoptosis and inhibit cell growth in Jurkat cell line.
Insights
Silencing the LSD1 gene in Jurkat cells inhibited proliferation and induced apoptosis. This suggests LSD1 is a potential therapeutic target for lymphocytic leukemia.
Area of Science:
- Molecular Biology
- Cell Biology
- Cancer Research
Background:
- The LSD1 gene plays a role in epigenetic regulation.
- Understanding LSD1's function is crucial for developing targeted cancer therapies.
Purpose of the Study:
- To investigate the impact of LSD1 gene silencing on Jurkat cell proliferation and apoptosis.
- To elucidate the underlying molecular mechanisms, including histone modifications.
Main Methods:
- RNA interference (RNAi) was used to silence the LSD1 gene in Jurkat cells.
- Quantitative reverse transcription PCR (RQ-PCR) and Western blotting assessed gene and protein expression.
- Cell proliferation was measured by MTT assay, and apoptosis was analyzed by flow cytometry.
Main Results:
- LSD1 gene silencing significantly reduced Jurkat cell proliferation and induced apoptosis.
- Apoptotic rates increased significantly in LSD1-silenced cells compared to controls.
- Downregulation of Bcl-2 and procaspase-3, and upregulation of Bax were observed, alongside altered histone methylation (H3K4me1, H3K4me2, Act-H3).
Conclusions:
- LSD1 gene depletion promotes apoptosis and inhibits cell growth in Jurkat cells.
- Histone methylation modifications are implicated in the observed effects.
- LSD1 may represent a viable therapeutic target for lymphocytic leukemia.
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