利用CRISPR/Cas9技术敲除LSD1基因显著抑制人慢性髓系白血病K562细胞的增殖与CD235a的表达

Knockout of LSD1 Gene by CRISPR/Cas9 System Significantly Inhibited Proliferation and Expression of CD235a in K562 Cells

目的:探索敲除LSD1基因对人慢性髓系白血病K562细胞产生的影响。方法:利用CRISPR/Cas9技术特异性敲除K562细胞系的LSD1基因;通过流式细胞分选技术获得单细胞后,经过扩增培养、Western blot与测序鉴定,分别获得1株LSD1^(+/-)与LSD1^(-/-)细胞株;MTS方法检测LSD1敲除对K562细胞增殖的影响;流式细胞分析技术检测LSD1对K562细胞表面标记表达的影响。结果:成功构建了K562的LSD1^(+/-)与LSD1^(-/-)细胞株;LSD1的敲除显著抑制了K562的增殖及CD235a的表达。结论:LSD1基因对K562细胞的增殖及CD235a的表达具有关键调控作用。

Objective： To study the effect of LSD1 knock-out on human chronic myeloid leukemia cells（K562 cells）.Methods： The LSD1 gene in K562 cells was knocked-out specifically by using CRISPR/Cas9 system,the single cells were gained by flow cytometric sorting technique,the LSD1 ^（＋/-）and LSD1^（-/-）cell lines were gained after amplificantion and culture,identification of Western blot and sequencing. The MTS assay was used to detect the effect of LSD1 knockout on the proliferation of K562 cells,the flow cytometry was used to examine the expression of K562 cell surface marker after LSD1 knockout. Results： The LSD1 stable knockout cell line of K562（LSD1 ^（＋/-）and LSD1^（-/-））were successfully costructed. It was found that knockout of LSD1 significantly inhibited the proliferation of K562 and the expression of CD235 a. Conclusion：LSD1 plays a key role in the regulation of K562 cell proliferation and CD235 a expression.