shRNA沉默LSD1基因对Jurkat细胞增殖的影响及机制研究

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™ 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. CONCLUSION: 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.