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目的 :了解和比较不同的反义c myc基因导入对HL 60细胞凋亡的影响。方法 :将分别载有c myc第 1、2和3外显子反义片段的三种重组逆转录病毒表达载体aM 1、aM 2和aM3导入体外培养的HL 60 ,并观察基因稳定表达对靶细胞凋亡的影响。结果 :(1 )光镜下aM2、aM 3组细胞核质比例明显缩小 ,出现凋亡小体等凋亡的形态特征 ;(2 )透射电镜下显示反义载体的导入使细胞胞质中出现了大量的自噬体和自噬泡 ,该现象的出现以aM1组为主 ;(3 )反义c myc导入引起HL 60中ACP酶活性增高 (aM1 >aM 3 >aM2 ) ,尤其是aM 1组 ,约提高了 86%。 (4 )流式细胞仪显示aM 2和aM3的瞬时和稳定表达都诱使HL 60凋亡率增高、细胞体积缩小、核质比减小 ,aM 1则无上述凋亡诱导作用。结论 :aM 2、aM3的导入促使HL 60出现了凋亡样改变 ,而aM 1则主要使细胞出现自噬。
OBJECTIVE: To understand and compare the effects of different antisense c myc gene on HL-60 cell apoptosis. Methods: Three recombinant retroviral vectors aM 1, aM 2 and aM3 containing the antisense fragments of exons 1, 2 and 3 of c myc were respectively introduced into HL 60 cells in vitro and stably expressed on target Effect of apoptosis. Results: (1) The proportion of apoptotic nuclei in aM2 and aM3 groups was significantly reduced under light microscope. (2) Transmission electron microscopy showed that the introduction of antisense vector resulted in the appearance of cytoplasm A large number of autophagosomes and autophagic vacuoles, the appearance of this phenomenon is mainly aM1 group; (3) The antisense c myc induces the increase of ACPase activity in HL 60 (aM1> aM3> aM2), especially the aM1 group , About an increase of 86%. (4) The transient and stable expression of aM 2 and aM3 induced the apoptosis rate of HL 60 induced by flow cytometry. The cell volume was reduced and the ratio of nucleus to cytoplasm was decreased. However, aM 1 had no apoptosis-inducing effect. CONCLUSION: The introduction of aM 2 and aM3 induces apoptosis-like changes in HL 60, while aM 1 mainly causes autophagy in cells.