The effects of shRNA-mediated interference on histone lysine specific demethylase 1 on cell proliferation and apoptosis in acute leukemia cell lines HL-60 and SHI-1

Abstract

Objective To investigate the effects of shRNA interference (RNAi) targeting the histone lysine specific demethylase 1 (LSD1) on the proliferation and apoptosis in acute leukemia (AL) cells. Methods LSD1 shRNA vectors were constructed and transduced into HL-60 and SHI-1 AL cell lines. Inhibitory efficiency of LSD1 was detected by real-time quantitative PCR (RT-qPCR) and Western blot respectively. Cell proliferation was assessed by methyl thiazolyl tetrazolium (MTT), and cell apoptosis was measured by flow cytometry. Results After interference of LSD1, the expression levels of LSD1 mRNA and protein in HL-60 and SHI-1 cells (mRNA: 0.242±0.023, 0.207±0.006; Protein: 0.256±0.015, 0.486±0.042) were decreased compared with blank control group without transfection process (mRNA: 1.021±0.178, 1.039±0.395; Protein: 0.552±0.026, 0.754±0.060) and empty vector negative control group (shNC group) (mRNA: 0.935±0.136, 1.016±0.203; Protein: 0.500±0.026, 0.750±0.049) (P < 0.05). The levels of their cell proliferation (absorbance value: 0.712±0.010, 0.549±0.007) were inhibited compared with blank control group (absorbance value: 0.823±0.010, 0.625±0.005) and shNC group(absorbance value: 0.818±0.019, 0.621±0.003) (P< 0.05). While cell apoptosis rates were increased [(32.80±1.35)%, (23.49±1.40)%] compared with blank control group [(8.08±0.62)%, (7.28±1.17)%] and shNC group[(8.08±0.62)%, (7.28±1.17)%] (P < 0.05). Conclusions Lentivirus-mediated shRNA interferencing LSD1 can inhibit cells' proliferation and promote apoptosis of HL-60 and SHI-1 AL cell lines, indicating that LSD1 may be a potential biological molecular marker and a new treatment target for AL. Key words: Leukemia, acute; Lysine specific demethylase 1; Cell proliferation; Apoptosis