腺病毒介导PDCD5基因转移促进依托泊甙诱导的K562细胞凋亡

本研究观察程序性细胞死亡5基因（programmed cell death5，PDCD5）重组腺病毒转染K562细胞后对化疗药物依托泊甙的增敏作用。利用AdMax^TM腺病毒载体包装系统，通过同源重组方法构建Ad—PDCD5重组腺病毒及对照腺病毒Ad-null及Ad-eGFP；用不同感染复数将Ad-eGFP、Ad—null或Ad—PDCD5转染人白血病细胞系，实时定量PCR检测PDCD5 mRNA的相对表达水平；利用MTT法及Annexin-V-FITC／PI双染色流式细胞术观察依托泊甙对转染后K562细胞增殖与凋亡的影响。结果表明：Ad-eGFP腺病毒对白血病细胞系K562、Jurkat及CEM的转染效率可达60％-86％。Ad—PDCD5重组腺病毒能梯度增加K562细胞PDCD5mRNA的相对表达水平，腺病毒介导的PDCD5基因转移促进依托泊甙诱导的K562细胞凋亡。结论：PDCD5重组腺病毒可能成为化疗药物的增敏刑。[第一段]

Adenovirus-mediated PDCD5 Gene Transfer Sensitizes Apoptosis of K562 Cells Induced by Etoposide

This study was purposed to investigate the effect of adenovirus-mediated transfer of PDCD5 gene on apoptosis of K562 cells induced by etoposide. Recombinant adenovirus PDCD5 (Ad-PDCD5), control vectors Ad-null and Ad-eGFP were constructed by AdMax vector system respectively. After K562 cells were transfected by Ad-PDCD5, Ad-null or Ad-eGFP with different multiplicity of infection (MOI), the expression level of the PDCD5 gene was examined by RQ-RT-PCR assay. The effects of etoposide in combination with Ad-PDCD5 on the proliferation and apoptosis of K562 cells were measured by using MTT assay and flow cytometry with Annexin-V-FITC/PI dual labeling technique, respectively. The results showed that the transfection efficiencies of Ad-eGFP in K562, Jurkat and CEM cells ranged from 60% to 86%. Expression level of PDCD5 gene in K562 cells was evidently increased following transfection with Ad-PDCD5. The Ad-PDCD5 synergistically enhanced the apoptotic percentage of K562 cells induced by VP-16, as compared with that of Ad-null + VP16 and VP-16 alone respectively. It is concluded that Ad-PDCD5 may be a potential agent for enhancing the chemotherapy effect.