摘要
目的探讨小干扰RNA（siRNA）沉默HOXA9基因对人类急性单核细胞白血病U937细胞株增殖、凋亡的影响。方法设计合成针对HOXA9的特异性siRNA寡核苷酸链，应用阳离子脂质体介导瞬时转染U937细胞。实验分为3组：实验组（脂质体转染靶向HOXA9的siRNA）、阴性对照组（脂质体转染阴性对照siRNA）和细胞对照组（仅加等量细胞及培养液）。利用反转录．聚合酶链反应法、Western blot法分别检测各组细胞HOXA9 mRNA、蛋白的表达；四甲基偶氮唑蓝（MTT）法检测细胞增殖，流式细胞术检测细胞凋亡。结果转染靶向HOXA9的siRNA后，实验组、阴性对照组、细胞对照组mRNA相对表达水平分别为、（22.980±0．548）％、（82．371±1．517）％、（8d．637±2．252）％（P〈0．05），蛋白相对表达水平分别为（50．377±2．773）％、（102．275±3．898）％、（107．510±3．905）％（P〈0．05）；siRNA转染后实验组U937细胞增殖明显受抑制且细胞凋亡率显著增高，24、48、72h增殖抑制率分别为（41．909±4．333）％、（54．470±3．756）％、（65．835±1．024）％，凋亡率为（26．800±2．081）％，与细胞对照组、阴性对照组比较差异均有统计学意义（P〈0．05）。结论靶向HOXA9的siRNA可有效沉默U937细胞HOXA9基因表达，明显抑制细胞的增殖并促进细胞凋亡，为临床白血病HOXA9基因靶向治疗提供了实验依据。
Objective To investigate the effects of small interference RNA （siRNA） targeting HOXA9 on the proliferation and apoptosis of human acute monocytic leukemia U937 cell line. Methods Effective and specific siRNA oligo targeting HOXA9 was designed and compounded. It was transfected transiently into U937 cells by cationic liposome. The cells was divided into three groups： experimental group（siRNA targeting HOXA9 was transfected by liposome）, negative control group （negative siRNA was transfected by liposome） and cell control group （add equal cells and medium）. The expression of HOXA9 mRNA and protein were detected by reverse transcription PCR and Western blot. The cell proliferation was assessed by MTY. The apoptosis of each group were measured by Annexin V-FITC. Results After transfected by siRNA targeting HOXA9, the relative mRNA expression levels of HOXA9 in the experimental group, negative control group and cell control group were （22.980±0.548） %, （82.371±1.517） % and （84.637±2.252） %, respectively （P 〈 0.05）, and the relative protein expression levels were （50.377±2.773）%, （105.500±3.900） % and （111.392±3.905） %, respectively （P 〈 0.05）. The inhibitory rates of cell proliferation and the apoptosis rates of the experimental group were significantly increased. The inhibitory rates of cell proliferation of 24 h, 48 h and 72 h were （41.909±4.333） %, （54.470±3.756） % and （65.835±1.024） %, respectively, and the apoatosis rate was （26.800±2.081） %. Compared with 2 controls, the experimental group differences had statistically significance （P 〈 0.05）. Conclusion siRNA targeting HOXA9 can effectively silence HOXA9 gene expression in U937 cell, suppress cell proliferation and induce cell apoptosis obviously, which providing experimental basis for clinical leukemia therapy by targeting HOXA9 gene.
出处
《肿瘤研究与临床》
CAS
2012年第8期533-536,共4页
Cancer Research and Clinic
基金
基金项目：山东省科学技术发展计划（2010GSF10264）
