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目的:研究RNA干涉(RNAi)Hsp701A基因的表达及其诱导慢性粒细胞白血病(慢粒)细胞株K562的凋亡。方法:构建Hsp701A基因小干涉RNA(siRNA)的真核表达载体,转染K562细胞并证实RNAi的有效性。用MTT比色法、AnnexinV-FITC/PI染色和流式细胞术,分别检测K562细胞的增殖、凋亡和细胞周期。结果:构建了Hsp701AsiRNA的真核表达载体。将其稳定转染K562细胞后,RNAi组细胞中Hsp701ARNA和蛋白的表达水平明显下降。Hsp701AsiRNA能抑制K562细胞增殖并诱导其细胞凋亡,将其阻滞于G1期。结论:RNAiHsp701A基因诱导的表达有望成为一种新的慢粒的治疗策略。
Objective: To study the expression of RNA interference (Hsp701A) gene and the apoptosis of K562 cells induced by chronic myelogenous leukemia (CML). Methods: Eukaryotic expression vector of small interfering RNA (siRNA) of Hsp701A gene was constructed and transfected into K562 cells to confirm the effectiveness of RNAi. The proliferation, apoptosis and cell cycle of K562 cells were detected by MTT assay, Annexin V-FITC / PI staining and flow cytometry. Results: The eukaryotic expression vector of Hsp701AsiRNA was constructed. After stable transfection of K562 cells, the expression level of Hsp701ARNA and protein in RNAi group decreased significantly. Hsp701AsiRNA can inhibit the proliferation of K562 cells and induce its apoptosis, blocking it in G1 phase. Conclusion: The expression induced by RNAiHsp701A gene is expected to be a new therapeutic strategy for CML.