Li Lei, Zhang Ai-Hua, Liu Ling-Bo, Bi Lan, Wang Li, Zhao Ya-Jie, Zou Ping
Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, Hubei Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2008 Apr;16(2):254-7.
This study was aimed to investigate the effect of small interfering RNA (siRNA) on the expression of mll-af9 oncogene and the proliferation of human acute monocytic leukemia cell line THP-1. One group of siRNA was designed targeting mll-af9 mRNA and finally obtained by chemosynthesis. Then the obtained siRNA was transfected into cultured human acute monocytic leukemia cell line THP-1 by lipofectamine. Flow cytometry was used to detect siRNA transfection efficiency. The level of mll-af9 mRNA expression was analyzed by reverse transcription polymerase chain reaction (RT-PCR). The cell proliferation rate was assayed by MTT. The change of cell cycles and apoptosis rate was detected by flow cytometry. The results showed that the siRNA transfection efficiency was 69.1%+/-1.8%. The level of mll-af9 mRNA expression was significantly inhibited in siRNA-transfected cells as compared with the controls. mll-af9-targeted siRNA inhibited the proliferation of THP-1 cells and induced cell apoptosis effectively after transfection. The percentage of G0/G1 phase cells significantly increased in siRNA-transfected cells in comparion with the control cells, but the percentage of S phase cells significantly decreased. It is concluded that the mll-af9-targeted siRNA can effectively inhibit the proliferation of human acute monocytic leukemia cell line THP-1.
本研究旨在探讨小干扰RNA（siRNA）对mll-af9癌基因表达及人急性单核细胞白血病细胞系THP-1增殖的影响。设计了一组靶向mll-af9 mRNA的siRNA，最终通过化学合成获得。然后将获得的siRNA用脂质体转染到培养的人急性单核细胞白血病细胞系THP-1中。采用流式细胞术检测siRNA转染效率。通过逆转录聚合酶链反应（RT-PCR）分析mll-af9 mRNA表达水平。采用MTT法检测细胞增殖率。通过流式细胞术检测细胞周期和凋亡率的变化。结果显示，siRNA转染效率为69.1%±1.8%。与对照组相比，siRNA转染细胞中mll-af9 mRNA表达水平显著受到抑制。靶向mll-af9的siRNA转染后有效抑制了THP-1细胞的增殖并诱导细胞凋亡。与对照细胞相比，siRNA转染细胞中G0/G1期细胞百分比显著增加，而S期细胞百分比显著降低。结论是，靶向mll-af9的siRNA可有效抑制人急性单核细胞白血病细胞系THP-1的增殖。
