Ma Xu-Dong, Zhao Ting, Huang Yi-Qun
Department of Hematology, Zhangzhou Affiliated Hospital of Fujian Medical University, Zhangzhou, Fujian Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2013 Feb;21(1):82-6. doi: 10.7534/j.issn.1009-2137.2013.01.018.
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(TM) 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 inhibits cell growth and induces cell apoptosis of KG-1 cell line, which may be a new therapeutic target in human leukemia.
本研究旨在探讨SUV39H1小干扰RNA（siRNA）对急性髓系白血病KG-1细胞系增殖和凋亡的影响。设计靶向SUV39H1基因的小干扰RNA（siRNA），并通过脂质体2000转染至KG-1细胞中。采用MTS法测定SUV39H1 siRNA对细胞生长的影响。通过流式细胞术检测细胞凋亡情况。采用蛋白质免疫印迹法检测P15以及抗凋亡蛋白如BCL-2、procaspase-9、procaspase-3和C-MYC的表达。结果表明，靶向SUV39H1的siRNA抑制了KG-1细胞的增殖。用30、60、120、240 nmol/L的SUV39H1 siRNA转染48 h后，增殖率分别为（76.43±1.98）%、（51.31±1.84）%、（37.31±1.61）%、（18.94±3.22）%，同时P15表达上调。凋亡细胞显著增加，用30、60、120 nmol/L靶向SUV39H1的siRNA转染后，凋亡率分别为（40.2±5.1）%、（56.8±4.8）%、（71.6±5.6）%（P<0.05）。转染后BCL-2、procaspase-9、procaspase-3、C-MYC的蛋白表达下调。结论是，靶向SUV39H1的siRNA抑制KG-1细胞系的细胞生长并诱导细胞凋亡，这可能是人类白血病的一个新的治疗靶点。
