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目的:观察c- myc基因反义寡核苷酸(ASODN)对HL- 60细胞增殖及c-myc基因表达的抑制作用,探讨 ASODN作为药物进行白血病基因治疗的可行性。方法:应用c -myc反义寡核苷酸封闭HL-60细胞c- myc基因,RT -PCR方法检测该基因表达抑制情况,流式细胞仪进行细胞周期分析,倒置显微镜观察细胞形态变化,绘制细胞生长 曲线,MTT法检测细胞增殖状态。结果:c- myc基因反义寡核苷酸转染HL -60细胞后,c -myc基因表达明显受抑;S 期细胞比值由55.6%降至16.7%;细胞生长受抑,增殖能力减弱,其抑制作用具有序列特异性及剂量依赖性。结 论:c -myc基因反义寡核苷酸对HL -60细胞增殖及c- myc基因的表达具有明显的抑制作用,有望成为抑制白血病细 胞增殖的核酸药物。
OBJECTIVE: To observe the inhibitory effect of c-myc antisense oligonucleotide (ASODN) on the proliferation and c-myc gene expression in HL-60 cells and to explore the feasibility of using ASODN as drug for gene therapy of leukemia. Methods: The c-myc gene of HL-60 cells was blocked by c-myc antisense oligonucleotide, the expression of c-myc gene was detected by RT-PCR, the cell cycle was analyzed by flow cytometry, the morphological changes of cells were observed by inverted microscope, Cell growth curve, MTT assay cell proliferation status. Results: After c-myc antisense oligonucleotide was transfected into HL-60 cells, the expression of c -myc gene was significantly inhibited; the cell ratio of S phase decreased from 55.6% to 16.7%; the cell growth was inhibited and the proliferation ability was weakened, Its inhibitory effect is sequence-specific and dose-dependent. CONCLUSION: Antisense c-myc antisense oligodeoxynucleotides can significantly inhibit the proliferation of HL-60 cells and the expression of c-myc gene, which is expected to become a nucleic acid drug that inhibits the proliferation of leukemia cells.