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目的 :研究 bcl- 2的反义寡核苷酸对足叶乙苷 (VP1 6 )诱导原代培养的急性白血病(AL)细胞凋亡的影响。方法 :用细胞计数法观察细胞的生存情况 ;用免疫荧光标记法通过流式细胞仪观测细胞 bcl- 2蛋白水平 ;用形态学观察及流式细胞仪检测细胞凋亡。结果 :靶向 bcl- 2 m RNA翻译起始区与靶向蛋白编码区的两个反义寡核苷酸分别与 VP1 6 联合作用 AL 细胞 4 8h,细胞的生存受到明显的抑制 ,分别同无关寡核苷酸 (NS- ODN)联合 VP1 6 组、单用 VP1 6 组进行比较 ,差异有显著性 (P<0 .0 5 )。这两个不同靶点的反义寡核苷酸分别与 VP1 6 联用 ,均能明显下调 AL 细胞 bcl- 2蛋白的表达 ,并且联合作用 AL 细胞 4 8h的凋亡细胞百分率分别与 NS- ODN联合 VP1 6 组、单用 VP1 6 组进行比较 ,差异有显著性 (P<0 .0 5 )。结论 :针对 bcl- 2 m RNA翻译起始区和蛋白编码区两个靶点的反义寡核苷酸能增强 VP1 6 诱导急性白血病细胞的凋亡。
AIM: To investigate the effect of antisense oligonucleotide bcl-2 on the apoptosis of primary cultured acute leukemia (AL) cells induced by etoposide (VP1 6). Methods: Cell survival was observed by cell counting method. Flow cytometry was used to observe the expression of bcl-2 protein by immunofluorescence staining. Cell apoptosis was detected by morphological observation and flow cytometry. RESULTS: Two antisense oligonucleotides targeting the translational initiation region of bcl-2 mRNA and the targeting protein coding region, respectively, inhibited the survival of AL cells 48 h after combined with VP1 6, and their survival was significantly inhibited Oligonucleotide (NS-ODN) combined with VP1 6 group, VP1 6 group alone, the difference was significant (P <0.05). The antisense oligonucleotides of these two different targets, respectively, combined with VP1 6, could significantly down-regulate the expression of bcl-2 protein in AL cells, and the percentage of apoptotic cells in the combined effect of AL cells 48 h and NS-ODN There were significant differences between VP1 6 group and VP1 6 group (P <0.05). CONCLUSION: Antisense oligonucleotides targeting both the translation initiation region and the protein coding region of bcl-2 mRNA enhance the apoptosis of acute leukemia cells induced by VP16.