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研究RNA干扰沉默HDAC1基因对髓系白血病HL-60细胞株细胞分化的影响。HDAC1基因的siRNA片段转染入HL-60细胞后,采用RT-PCR、Western blotting分别检测HDAC1 mRNA和蛋白的表达变化,Wright-Giemsa染色观察细胞形态,流式细胞术分析CD13、CD33、CD14的表达变化,NBT还原比色实验观察细胞分化能力。结果表明,HDAC1 siRNA可沉默该基因表达;HDAC1 siRNA的浓度为30～60 nmol·L-1时,24 h后细胞形态向成熟粒系分化;60 nmol·L-1组的NBT还原能力为0.25±0.02,与对照组比较差异有统计学意义(P<0.05);但更高浓度组却无明显变化;60 nmol·L-1组细胞CD13、CD33表达率分别为(96.50±0.70)%、(66.73±0.50)%,而对照组分别为(3.39±0.68)%、(96.80±1.70)%,差异有统计学意义(P 0.000 5);但是CD14的表达率无明显变化。结果提示,HDAC1 siRNA的浓度在30～60 nmol·L-1时可诱导细胞向成熟粒细胞分化,有望成为白血病靶向基因治疗的新工具。
To study the effect of HDAC1 silencing RNA interference on the cell differentiation of HL-60 myeloid leukemia cells. The expression of HDAC1 mRNA and protein were detected by RT-PCR and Western blotting respectively after transfection of siRNA fragment of HDAC1 gene into HL-60 cells. The cell morphology was observed by Wright-Giemsa staining. The expressions of CD13, CD33 and CD14 were analyzed by flow cytometry Expression change, NBT reduction colorimetric assay to observe cell differentiation ability. The results showed that HDAC1 siRNA silenced the expression of this gene. When the concentration of HDAC1 siRNA was 30 ~ 60 nmol·L-1, the cell morphology was differentiated into mature granulocyte line after 24 h. The NBT reduction capacity of 60 nmol·L-1 group was 0.25 (P <0.05), but there was no significant change in the higher concentration group. The expression rates of CD13 and CD33 in 60 nmol·L-1 group were (96.50 ± 0.70)%, (66.73 ± 0.50)%, while the control group were (3.39 ± 0.68)% and (96.80 ± 1.70)% respectively, the difference was statistically significant (P 0.000 5). However, the expression of CD14 had no significant change. The results suggest that the concentration of HDAC1 siRNA at 30 ~ 60 nmol·L-1 can induce cells to differentiate into mature granulocytes and is expected to become a new tool for targeted gene therapy of leukemia.