三种反义c-myc RNA对HL-60细胞凋亡的影响

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。

EFFECTS OF THREE KINDS OF ANTISENSE C-MYC RNA ON THE APOPTOSIS OF HL-60 CELLS

Objective:To investigate the effects of different antisense c myc molecule on the apoptosis of HL 60.Methods:Three antisense retrovirus expression vectors named aM1,aM2 and aM3 for human c myc exon 1,2 and 3 respectively,were tranfered into cultured HL 60 cells,then stable expression assay were carried out to study their effects on target cells.Results:(1)Under microscope,the nuclear/plasma ratio of cells in aM2 and aM3 groups were reduced,and morphological signs of apoptosis,such as apoptosis body,appeared;(2)Flow cytometric analysis showed that both transient and stable expressions of aM2 and aM3 could induce the apoptosis,minified cell size and the muclear/plasma ratio.Nevertheless,aM1 had little effect;(3)Electronic microscope examination showed that numerous autophagic vacuoles and autophagosomes occurred in the cells transfected with antisense c myc,especially with aM1;(4)ACP activity were improved by antisense c myc (aM1>aM3>aM2),especially in aM1 group(increased about 86%).Conclusion:aM2 and aM3 may induce HL 60 cells apoptosis,while aM1 mainly induced them into the autophagy pathway.