关键词:鸟氨酸脱羧酶;寡核苷酸类,反义;白血病,幼红细胞,急性
摘要 目的: 观察鸟氨酸脱羧酶反义寡核苷酸(ODCasODN)引起鸟氨酸脱羧酶(ODC)表达及活性的抑制对人红白血病K 562细胞生长、凋亡的影响,为探索抑制多胺生物合成的抗癌新途径提供实验依据。方法: 采用硫代修饰的ODCasODN处理细胞,观察生长曲线,及用Giemsa染色观察形态变化,RNA斑点杂交检测基因表达,放射微量法测定ODC活性。结果: ODCasODN能抑制K 562细胞生长、诱导凋亡,并证明了该细胞的ODC、Bcl-2表达下降及ODC活性受到抑制。结论: ODCasODN能下调ODC表达、降低其酶活性,引起bcl-2基因表达抑制,导致K 562细胞生长抑制及凋亡诱导。
EFFECT OF ODCasODN ON APOPTOSIS OF HUMAN ERYTHROLEUKEMIA
CELL K 562 AND ITS ASSOCIATED GENE EXPRESSION
Xiang Qing,Cao Shulan,Xu Mei,et al
(Department of Biochemistry,Institute of Clinical Medical Sciences)
Abstract Objective: To observe the inhibition of ornithine decarboxylase antisense oligodeoxynucleotide(ODCasODN) on expression and activity of ornithine decarboxylase(ODC) and its effect on the growth and apoptosis of human erythroleukemia cell K 562 for searching the new approach of anticancer by inhibition of polyamine biosynthesis.Methods : The cellular morphology and growth rate of K 562 treated with ODC antisense phosphoro thioate ODN were observed by using Giemsa sataining and cell counting.The detection of associated gene expression and determination of ODC activity were performed by using RNA dot-blotting and microradioanalysis respectively.Results : ODCasODN inhibited the growth of K 562 cell,induced apoptosis and caused the downregulation of ODC and bcl-2 expression.Conclusion : The inhibition of polyamine biosynthesis which results from the suppression of ODC expression by ODCasODN can downregulate the expression of bcl-2 gene and elicits growth inhibition and apoptosis induction of the treated cells thereby.
Subject headings : ornithine decarboxylase;oligonucleotides,antisense;leukemia,erythroblastic,acute
生理性多胺包括腐胺、精脒及精胺 ......
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