Effects of siRNA Targeting hoxa9 and Low Dose of Cytosine Arabinoside on Proliferation and Apoptosis of U937 Cells
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摘要: 目的 探讨利用小干扰RNA（small interference RNA，siRNA）沉默hoxa9基因联合小剂量阿糖胞苷（Ara-C）对U937细胞增殖、凋亡的影响。方法设计合成针对hoxa9的特异性siRNA，应用脂质体介导转染U937细胞。利用RT-PCR法、Western blot法分别检测各组细胞hoxa9 mRNA、蛋白的表达；siRNA和（或）小剂量Ara-C分别作用U937细胞后，应用MTT法检测细胞增殖抑制率；流式细胞术检测细胞凋亡率。结果靶向hoxa9的siRNA可有效沉默hoxa9的表达，hoxa9 mRNA及蛋白相对水平明显下降。RNAi联合小剂量Ara-C作用于U937细胞后对细胞增殖抑制作用最强，48 h细胞凋亡率显著升高，与其余组比较均有统计学差异（P<0.05）。结论靶向hoxa9的siRNA联合小剂量Ara-C能显著抑制U937细胞增殖并促进其凋亡，为研究hoxa9联合化疗治疗白血病的作用机制及应用于临床提供实验依据。Abstract: Objective To investigate the effect of small interference RNA(siRNA)targeting hoxa9 and low dose of cytosine arabinoside(Ara-C)on the proliferaion 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 into U937 cells by liposome.The expression of hoxa9 mRNA and protein were detected by reverse transcription PCR and Western blot.The effects of siRNA targeting hoxa9 and(or) low dose of Ara-C on the cell viability of U937 were detected by MTT assay.The apoptosis rate in each group was measured by Annexin V-FITC. Results The expression of hoxa9 was effectively silence by siRNA targeting hoxa9.As a result,the mRNA expression of hoxa9 and the protein decreased obviously.The inhibitory rate(IR)of cell proliferation in group “RNAi+low dose of Ara-C” was the most significant.And the apoptosis rate at 48 h in this group increased apparently,which was significantly different with those in the other groups(P<0.05). Conclusion siRNA targeting hoxa9 combined low dose of Ara-C can significantly inhibit the proliferation of U937 cells and promote its apoptosis.This work offers an experimental data for revealing the interaction mechanism and for the clinic applications of hoxa9 combined with chemotherapy in the treatment of leakemia.
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Key words:
- RNA interference /
- Cytosine arabinoside /
- HomeoboxA9 /
- U937 cells /
- Leukemia
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