沉默SUV39H1基因对急性髓系白血病KG-1细胞凋亡的影响

摘要

本研究旨在探讨沉默SUV39H1基因对急性髓系白血病细胞株KG-1增殖和凋亡的影响.将SUV39H1 siRNA经LipofectamineTM2000转染至KG-1细胞,应用MTS法检测细胞增殖率,观察SUV39H1 siRNA对KG-1细胞增殖的影响；流式细胞术分析细胞凋亡；Western blot检测SUV39H1 siRNA作用后P15和凋亡相关蛋白BCL-2、prcaspase-9、procaspase-3、C-MYC的表达.结果表明,沉默SUV39H1基因可抑制细胞增殖,SUV39H1 siRNA浓度为30、60、120、240 nmol/L作用48 h后,KG-1细胞的增殖率分别为(76.43±1.98)％、(51.31±1.84)％、(37.31±1.61)％、(18.94±3.22)％,差异具有统计学显著意义(P＜0.05)；SUV39H1 siRNA可上调P15基因的表达；SUV39H1 siRNA可诱导细胞凋亡,30、60、120 nmol/L SUV39H1 siRNA作用48 h后,KG-1细胞凋亡率分别为(40.2±5.1)％、(56.8±4.8)％、(71.6±5.6)％,差异有统计学显著意义(P＜0.05)；抗凋亡相关蛋白BCL-2、prcaspase-9、procaspase-3、C-MYC的表达减少.结论:SUV39H1 siRNA能抑制KG-1细胞的增殖并诱导其凋亡,有望成为白血病治疗的一个新的靶点.%This study was aimed to investigate the effects of SUV39H1 siRNA on proliferation and apoptosis of acute myelogenous leukemia KG-1 cell line. The small interfering RNA( siRNA) targeting SUV39H1 gene was designed and transfected into KG-1 cells by Lipofectamine?2000. Cell growth affected by SUV39H1 siRNA was determined by MTS method. Cell apoptosis was measured by flow cytometry. The expressions of P15 and anti-apoptosis protein such as BCL-2, procaspase-9, procaspase-3 and C-MYC were detected by Western blot. The results indicated that siRNA targeting SUV39H1 inhibited proliferation of KG-1 cells. Proliferated rates were (76.43 ± 1.98) % , (51.31 ±1.84)% , (37.31 ±1.61)% , (18.94 ± 3. 22) % respectively after transfection with SUV39H1 siRNA at 30, 60, 120, 240 nmol/L for 48 h, while P15 expression was upregulated. Apoptotic cells significantly increased, apoptotic rates were (40. 2 ± 5.1)% , (56.8±4.8)% , (71.6 ±5.6)% respectively after transfection with siRNA targeting SUV39H1 at 30, 60, 120 nmol/L(P<0.05). The protein expression of BCL-2, procaspase-9, procaspase-3, C-MYC was downregulated after transfection. It is concluded that the siRNA targeting SUV39H1 inhibites cell growth and induces cell apoptosis of KG-1 cell line, which may be a new therapeutic target in human leukemia.