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本研究观察程序性细胞死亡5基因(programmed cell death 5,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AdMaxTM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PDCD5mRNA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PDCD5 mRNA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剂。
This study was to investigate the sensitizing effect of chemotherapeutic etoposide after transfection of K562 cells with the programmed cell death 5 (PDCD5) recombinant adenovirus. Using AdMaxTM adenovirus vector packaging system, Ad-PDCD5 recombinant adenovirus and control adenovirus Ad-null and Ad-eGFP were constructed by homologous recombination method. Ad-eGFP, Ad-null or Ad-PDCD5 Human leukemia cell line was used to detect the relative expression level of PDCD5 mRNA by real-time quantitative PCR. The effect of etoposide on the proliferation and apoptosis of K562 cells was observed by MTT assay and Annexin-V-FITC / PI double staining flow cytometry. The results showed that Ad-eGFP adenovirus transfected leukemia cell lines K562, Jurkat and CEM up to 60% -86% efficiency. Ad-PDCD5 recombinant adenovirus can increase the relative expression of PDCD5 mRNA in K562 cells. Adenovirus-mediated PDCD5 gene transfer promotes etoposide-induced K562 cell apoptosis. Conclusion: PDCD5 recombinant adenovirus may become a sensitizer for chemotherapeutic drugs.