目的:研究小干扰RNA(RNAi)技术沉默凋亡抑制蛋白Apollon基因联合小剂量阿糖胞苷(Ara-C)对K562细胞化疗敏感性的影响。方法:构建pGPHI-GFP-Neo-Apollon真核表达载体,经阳离子脂质体转染K562细胞,实验分为RNAi组、阴性质粒组、细胞对照组、Ara-C组和RNAi+Ara-C组5组,采用RT-PCR和细胞免疫荧光检测干扰效果。RNAi与小剂量Ara-C单独和联合应用后,采用MTT法检测细胞增殖能力,流式细胞术检测细胞凋亡率。结果:RT-PCR结果显示,实验组Apollon mRNA的相对表达量为(27.622±0.057)%,与细胞对照组(89.770±0.028)%和阴性对照组(84.868±0.057)%相比明显降低,差异有统计学意义,F=57.573,P=0.012。细胞免疫荧光检测结果显示,实验组细胞Apollon蛋白的荧光强度为(15.96±1.71)%,明显低于细胞对照组(35.36±2.90)%和阴性对照组(34.97±2.65)%,差异有统计学意义,F=71.063,P=0.013。重组载体和(或)10μg/mL Ara-C作用于K562细胞24、48和72h后,RNAi组和RNAi+Ara-C组K562细胞增殖抑制率(IR%)明显增高,随着作用时间的延长,抑制效果越明显。流式细胞术结果显示,RNAi+Ara-C组细胞凋亡率为(30.816±1.06)%,明显高于细胞对照组(4.803±0.112)%、阴性对照组(4.566±0.205)%和Ara-C组(11.61±0.604)%及RNAi组(20.226±0.986)%,差异有统计学意义,F=138.57,P<0.001。结论:靶向Apollon RNAi联合小剂量Ara-C能有效抑制K562细胞增殖并促进其凋亡,显著减少Ara-C使用剂量,明显提高了K562细胞对Ara-C的敏感性。 Objective: To study the effect of small interfering RNA (RNAi) technology on the chemosensitivity of K562 cells induced by Apollon gene combined with small dose of Ara-C. Methods: The eukaryotic expression vector pGPHI-GFP-Neo-Apollon was constructed and transfected into K562 cells by cationic liposome. The experiment was divided into RNAi group, negative plasmid group, cell control group, Ara-C group and RNAi + Ara-C group 5 groups. The interference effect was detected by RT-PCR and immunofluorescence. RNAi and low-dose Ara-C alone and in combination, the use of MTT assay of cell proliferation, flow cytometry detection of apoptosis rate. Results: RT-PCR results showed that the relative expression of Apollon mRNA in experimental group was (27.622 ± 0.057)%, which was significantly lower than that in cell control group (89.770 ± 0.028)% and negative control group (84.868 ± 0.057)%, Statistically significant, F = 57.573, P = 0.012. The results of immunofluorescence showed that the fluorescence intensity of Apollon protein in experimental group was (15.96 ± 1.71)%, which was significantly lower than that in cell control group (35.36 ± 2.90)% and negative control group (34.97 ± 2.65)%, the difference was statistically significant Meaning, F = 71.063, P = 0.013. After treated with recombinant vector and / or Ara-C at 10μg / mL for 24,48 and 72h, the proliferation inhibition rates (IR%) of K562 cells in RNAi group and RNAi + Ara-C group were significantly increased. With the prolongation of action time , The more obvious the inhibitory effect. The results of flow cytometry showed that the rate of apoptosis in RNAi + Ara-C group was (30.816 ± 1.06)%, which was significantly higher than that in cell control group (4.803 ± 0.112)%, negative control group (4.566 ± 0.205)% and Ara- C group (11.61 ± 0.604)% and RNAi group (20.226 ± 0.986)%, the difference was statistically significant, F = 138.57, P <0.001. Conclusion: Targeting Apollon RNAi combined with low-dose Ara-C can effectively inhibit the proliferation and promote the apoptosis of K562 cells, significantly reduce the dose of Ara-C and significantly increase the sensitivity of K562 cells to Ara-C.