小分子干扰RNA抑制HLX1表达对急性髓系白血病细胞侵袭的影响及机制探讨

Inhibitory effect of HLX1 on proliferation and invasion of acute myeloid leukemia cell

目的:探讨小分子干扰RNA(siRNA)抑制人同源异型盒基因(H2.0-like homeobox,HLX1)表达对急性髓系白血病细胞系U937细胞增殖与侵袭能力的影响。方法:利用HLX1特异的siRNA瞬时转染U937细胞,通过实时定量(RTPCR)与蛋白印迹法(Western blot)分别测定HLX1的mRNA及蛋白质的表达水平,应用细胞体外增殖实验法(MTT)与体外侵袭实验观察siRNA抑制HLX1表达后的U937细胞增殖与侵袭变化。结果:相对于空白对照组,HLX1特异的siRNA转染24 h后可抑制U937细胞HLX1 mRNA表达,抑制率为(70.0±2.7)%(P<0.01);转染48 h后可有效抑制HLX1蛋白质表达,抑制率为(81.0±3.1)%(P<0.01);转染组U937细胞增殖能力显著下降(P<0.01),转染组穿过基质胶的U937细胞数(22.0±2.3)显著低于空白对照组(66.0±5.0)(P<0.01),p-21活化激酶1(PAK1)蛋白表达水平显著下降(P<0.01)。结论:siRNA下调HLX1表达水平可显著抑制急性髓系白血病细胞的增殖与侵袭能力。

AIM： To explore the effect of HLX1 on the proliferation and invasion ability in acute myeloid leukemia cell U937. METHODS： The HLX1mRNA-targeting siRNA was transfected to U937 cells,the mRNA and protein expression of HLX1 were assessed by RT-PCR and Western blot,respectively. MTT assay and Transwell chamber assay were used to observe the proliferation and invasion ability of U937 cell after being transfected with HLX1 mRNA-targeting siRNA. RESULTS： Compared with the control group,HLX1 siRNA transfection reduced the level of HLX1 mRNA in U937 cells after 24 h,the inhibition rate was（ 70. 0 ± 2. 7） %（ P 0. 01）; HLX1 siRNA transfection also reduced the expression of HLX1 protein in U937 cells after 48 h,the inhibition rate was（ 81. 0 ± 3. 1） %（ P 0. 01）; HLX1 transfection group cell proliferation was inhibited significantly（ P 0. 01）; The invasion cells of HLX1 siRNA（22. 0 ± 2. 3） were significantly lower than that of control group（66. 0 ± 5. 0）（P 0. 01）,the expression of PAK1 protein was decreased in HLX1 transfection group（ P 0. 01）.CONCLUSION： SiRNA can significantly inhibit the proliferation and invasion of acute myeloid leukemia cell by down-regulating the expression of HLX1.