抑癌基因Hes1影响急性髓系白血病细胞增殖和凋亡的机制研究

Inhibitory effects of Hes1 on acute myeloid leukemia cells

目的 阐明Hes1与急性髓系白血病（AML）细胞增殖和凋亡的关系.方法 通过实时定量PCR检测AML原代细胞和HL-60、U937、KG1a细胞中Hes1和p21的表达情况;通过在AML细胞中转染逆转录病毒载体使Hes1高表达,通过MTT及流式细胞术检测高表达Hes1的AML细胞增殖和细胞周期、凋亡的改变;并通过成瘤实验检测Hes1＋AML细胞在NOD/SCID小鼠体内的增殖情况.结果 Hes1和p21在AML患者原代细胞和HL-60、U937、KG1a细胞中的表达分别为0.67±0.24和0.59±0.43、0.42±0.03和0.32±0.26、0.54±0.01和0.44±0.12、0.36±0.12和0.59±0.43,均较正常对照组水平降低（P值均＜0.05）;通过逆转录病毒载体诱导后HL-60、U937、KG1a细胞中Hes1的表达分别为4.9±0.2、5.2±0.4、5.8±0.5,均较未转染诱导前上调（P值均＜0.05）;感染Hes1后AML细胞与感染空载体的AML细胞比较,增殖受到抑制,细胞凋亡增加.与对照组比较,3种细胞系高表达Hes1后在NOD/SCID小鼠体内的成瘤性均降低（P值均＜0.05）.结论 Hes1过表达可抑制AML细胞的增殖,诱导AML细胞凋亡,从而提示Hes1为AML的抑制基因,可能成为治疗AML的新靶点.

Objective To elucidate the impact of Hesl on the proliferation and apoptosis of acute myeloid leukemia （AML） cells.Methods The expression levels of Hesl and p21 in AML patient samples and myeloid leukemia cell lines were analyzed by real-time PCR.Hes1 was up-regulated by retrovirus transfection in AML cell lines and the proliferation capacity were assayed by MTT,cell cycle by Hoechst/PY,apoptosis by AnnexinV.Results The expression of Hes1 in primary AML cells and HL-60、U937 、KG1a cell lines were 0.67±0.24,0.59±0.43,0.42±0.03,and 0.32±0.26,respectively,and p21 were 0.54±0.01,0.44±0.12,0.36±0.12,and 0.59±0.43,respectively.Hes1 expression levels after transduction in HL-60、U937、KG1a were 4.9±0.2,5.2±0.4,5.8±0.5,respectively.Induced activation of Hesl led to AML cells growth arrest and apoptosis,which was associated with an enhanced p21 expression.Besides,activated Hes1 led to AML cells growth inhibition in vivo.Conclusions Hesl could mediate growth arrest and apoptosis in AML cells,which may be a novel target for AML.