RNA干扰PCGF4/Bmi-1抑制白血病K562细胞系增殖

Overexpression of PCGF4 /Bmi-1 gene suppressed by RNA interference reduces the proliferative rate of leukemia K562 cell line

目的:利用RNAi方法针对PCGF4/Bmi-1基因,抑制其在慢性髓性白血病K562细胞中的过表达,观察K562细胞的增殖性改变。方法:RT-PCR检测PCGF4/Bmi-1基因在K562中的表达情况。利用Invitrogen公司RNAi慢病毒表达载体系统,构建针对PCGF4/Bmi-1基因慢病毒RNAi表达载体。转染K562细胞,瞬时干扰PCGF4/Bmi-1基因的表达,实时定量PCR检测干扰前后基因表达变化。利用生长曲线测定干扰载体转染前后细胞生长速度变化。结果:PCGF4/Bmi-1慢病毒RNAi表达载体成功地构建,并建立瞬时干扰PCGF4/Bmi-1基因的K562细胞系。实时定量PCR检测,PCGF4/Bmi-1基因在K562细胞系中过表达被显著抑制。生长曲线测定,PCGF4/Bmi-1基因干扰后细胞增殖明显变慢。结论:干扰PCGF4/Bmi-1基因,降低该基因表达后能明显减低细胞的生长速度。

Objective： To explore the effect of suppressing PCGF4/Bmi-1 overexpression in CML cells line K562, on the proliferative rate of K562 cells . Methods： RT-PCR was used to detect the expression of PCGF4/Bmi-1 gene in K562 cells. BLOCK-iT^TM Lentiviral RNAi expression vector bought from Invitrogen Company on PCGF4/Bmi-1 was constructed. After transient transfection,into K562 cells, real-time quantitation PCR was used to detect the expression alteration of PCGF4/Bmi-1, and growth curve was utilized to observe the alteration of growth rate of cells. Results： PCGF4/Bmi-1 BLOCK-iTTM Lentiviral RNAi expression vector was successfully constructed and K562 cells of transient interfering PCGF4/Bmi-1 were built. Overexpression of PCGF4/Bmi-1 gene was greatly suppressed by detection of real-time quantitation PCR, and Proliferative rate of K562 cells was markedly decreased using growth curve detection after transient transfection of PCGF4/Bmi-1 BLOCK-iTTM Lentiviral RNAi expression vector into K562 cells. Conclusion： Overexpression of PCGF4 /Bmi-1 gene suppressed by RNA Interference reduces the proliferative rate of leukemia K562 cell line.