转染p16与dll4阻滞白血病细胞株K562细胞周期

Arresting Effect of p16 and dll4 Transfection on Cell Cycle of K562 Cells

本研究探讨p16,dll4基因真核表达载体在白血病细胞中的表达和作用,设计并构建包含野生型p16cDNA,dll4cDNA的真核双表达载体pBudCE4.1-16-dll4,用脂质体法转染白血病细胞,用Western blot方法检测外源性p16及dll4的表达情况;通过CCK-8和流式细胞仪检测细胞的生长曲线和周期。结果表明:成功构建p16、dll4基因双表达真核载体,体外成功转染入白血病细胞。在白血病细胞检测到外源性P16、Dll4蛋白的表达。转染48小时后,试验组与对照组比较,细胞周期被阻滞(p<0.001),细胞增殖下降(p<0.001)。结论:重组质粒pBudCE4.1-16-dll4体外转染白血病细胞后,两个目的基因能够在细胞中同时表达,并诱发细胞周期阻滞于G0/G1期。

This study was purposed to investigate the expression and role of eukaryotic expression vector containing pl6, dl14 genes in leukemia K562 ceils. A vector pBudCFA. 1-16-d114 containing wild type pl6cDNA and dlI4cDNA was designed and constructed, then this vector was transfected into leukemia K562 cells by using lipofectamine 2000. The expression of pl6 and dl14 genes was detected by Western blot, the cell growth curve and cell cycle were determined by CCK-8 kit and flow cytometry respectively. The results showed that the recombinant plasmid pBudCE4.1-16-d114 was constructed and transfected into I（562 cells in vitro successfully. The expression of exogenous P16 and DII4 proteins could be detected in K562 cells. After transfection for 48 hours, the K562 cells were arrested in G1 phase, the cell count increased in G0/G1 phase and reduced in S phase, the cell proliferation decreased as compared with control. It is concluded that the pl6 and dl14 genes can simultaneously express in K562 cells transfected with recombinant plasmid pBudCE4. 1-16 -dl14 in vitro which results in G0/G1 arrest and reduces cell proliferation.