Design of antisense oligodeoxynucleotide targeting at bcl-2 mRNA and observation on its effect on the sensitivity of leukemia cells to arabinosyl cytosine.
- Author:
Xiao-Yong LEI
1
;
Yuan ZHANG
Author Information
1. Institute of Hematology, Medical College of Jinan University, Guangzhou, 510632, China.
- Publication Type:Journal Article
- MeSH:
Antimetabolites, Antineoplastic;
pharmacology;
Apoptosis;
drug effects;
Base Sequence;
Cell Division;
drug effects;
Cell Survival;
drug effects;
Cytarabine;
pharmacology;
Drug Design;
Drug Synergism;
HL-60 Cells;
Humans;
Inhibitory Concentration 50;
K562 Cells;
Leukemia;
drug therapy;
pathology;
Molecular Sequence Data;
Nucleic Acid Conformation;
Oligonucleotides, Antisense;
genetics;
pharmacology;
Proto-Oncogene Proteins c-bcl-2;
genetics;
metabolism;
RNA, Messenger;
chemistry;
genetics
- From:
Journal of Experimental Hematology
2003;11(1):41-44
- CountryChina
- Language:Chinese
-
Abstract:
The effective target sites for antisense oligodeoxynucleotides (AS-ODN) on bcl-2 mRNA, except its start region, were looked for and their effects on the sensitivity of HL-60 and K562 leukemia cells to arabinosyl cytosine (Ara-C) were observed. The secondary structure of bcl-2 mRNA was simulated with RNAstructure microsoftware, and AS-ODNs, targeting at some sites, were designed and synthesizd with phosphorothioate modifying. The median inhibitory concentration (IC(50)) of Ara-C for HL-60 and K562 cells was determined with MTT method; the apoptosis rate and bcl-2 protein expression level were assayed by flow cytometry. The results showed that 10 micro mol/L bcl-2 AS-ODN combined with Ara-C inhibited the expression of bcl-2 protein, increased apoptosis in HL-60 and K562 cells and decresed IC(50) of Ara-C significantly. The AS-ODN targeting the coding region of bcl-2 mRNA had stronger effect than AS-ODN targeting the translation initiation region. In conclusion, the much more effective sites for antisense oligodeoxynucleotides to target, except translation start region, might provide a new useful way for antisense drug design.
