干扰组蛋白赖氨酸特异性去甲基化酶1基因对急性白血病HL-60和SHI-1细胞增殖及凋亡的影响

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

目的：探讨shRNA干扰组蛋白赖氨酸特异性去甲基化酶1（LSD1）基因对急性白血病（AL）细胞增殖与凋亡的影响。方法构建慢病毒载体介导LSD1干扰的AL稳定细胞株HL-60和SHI-1，采用实时荧光定量聚合酶链反应（PCR）和Western blot方法检测两细胞株LSD1抑制效果，通过四甲基偶氮唑盐（MTT）法检测细胞增殖，采用流式细胞术检测细胞凋亡。结果 LSD1干扰后，HL-60和SHI-1细胞株的LSD1 mRNA和蛋白表达水平（mRNA：0.242±0.023、0.207±0.006，蛋白：0.256±0.015、0.486±0.042）较未经转染处理的空白对照组（mRNA：1.021±0.178、1.039±0.395，蛋白：0.552±0.026、0.754±0.060）和空载体阴性对照组（shNC组）（mRNA：0.935±0.136、1.016±0.203，蛋白：0.500±0.026、0.750±0.049）均下调（P＜0.05），且细胞增殖水平（吸光度值分别为0.712±0.010、0.549±0.007）低于空白对照组（吸光度值分别为0.823±0.010、0.625±0.005）和shNC组（吸光度值分别为0.818±0.019、0.621±0.003）（P＜0.05），而细胞凋亡率[（32.80±1.35）%、（23.49±1.40）%]高于空白对照组[（8.08±0.62）%、（7.28±1.17）%]和shNC组[（8.00±0.32）%、（7.19±0.65）%]（P＜0.05）。结论慢病毒载体介导的shRNA干扰LSD1使AL细胞株HL-60和SHI-1增殖受抑制，凋亡增加。 LSD1有可能成为AL的生物分子标志和治疗新靶点。

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.